Multi-omics analysis reveals the influence of tetracycline on the growth of ryegrass root.

Environmental factors, including antibiotics such as tetracycline, can alter biological processes in plants. To ascertain how cell/tissue response to tetracycline, a multi-omic analysis was implemented to explore the molecular mechanism of tetracycline influencing the growth of ryegrass root. Tetracycline induced extensive changes in the root metabolome in plants, particularly impacting metabolites of flavonoid metabolic pathways, which were supported through consistent differences between transcriptome and proteome. Cross-comparison between mRNA and protein contents considered the authentication of congruence with related metabolites and revealed changes of several biological processes under tetracycline stress. Overall, we present an undemanding multi-omic strategy to survey the significant influence on the root under tetracycline stress.

Is negative density-dependent reproduction regulated by density-induced stress in root voles? Two field experiments.

Density dependence in reproduction plays an important role in stabilizing population dynamics via immediate negative feedback from population density to reproductive output. Although previous studies have shown that negative density-dependent reproduction is associated with strong spacing behavior and social interaction between individuals, the proximal mechanism for generating negative density-dependent reproduction remains unclear. In this study, we investigated the effects of density-induced stress on reproduction in root voles. Enclosed founder populations were established by introducing 6 (low density) and 30 (high density) adults per sex into per enclosure (four enclosures per density in total) during the breeding season from April to July 2012 and from May to August 2015. Fecal corticosterone metabolite (FCM) levels, reproductive traits (recruitment rate and the proportion of reproductively active individuals), and founder population numbers were measured following repeated live trapping in both years. The number of founders was negatively associated with recruitment rates and the proportion of reproductively active individuals, displaying a negative density-dependent reproduction. FCM level was positively associated with the number of founders. The number of founder females directly affected the proportion of reproductive females, and directly and indirectly through their FCM levels affected the recruitment rate; the effect of the number of male founders on the proportion of reproductive males was mediated by their FCM level. Our results showed that density-induced stress negatively affected reproductive traits and that density-induced stress is one ecological factor generating negative density-dependent reproduction.

First genotyping of Enterocytozoon bieneusi in plateau pikas (Ochotona curzoniae) from the Qinghai Plateau, Northwest China.

Enterocytozoon bieneusi is considered the most common microsporidian species and is frequently detected in humans and various animals worldwide. However, information on E. bieneusi infection in plateau pikas (Ochotona curzoniae) is somewhat limited. Therefore, this study was conducted to determine the infection status and genotype characteristics of E. bieneusi in plateau pikas in China. A total of 33 fresh fecal samples were collected from plateau pikas captured on the Qinghai Plateau. By PCR assay and DNA sequencing of the ITS gene, 5 (15.2%, 5/33) isolates were diagnosed as positive. Sequence analysis revealed the presence of one known sheep-derived genotype, CHS17 (4/5), and a novel E. bieneusi genotype, CHPP1 (1/5), with maximum shared identity with the CHS17 genotype. Phylogenetically, these isolates were clustered into subgroup 1i of zoonotic group 1 with genotypes CHS17 and CHN14 from plateau ruminants, but the new CHP1 genotype formed a cluster separate from those genotypes. This is the first report of E. bieneusi in plateau pikas. The findings suggested that these animals might be potential reservoirs of zoonotic E. bieneusi, and transmission of the pathogen between plateau pikas and sheep probably occurs in the region.

Ghrelin attenuates infectious bursal disease virus-induced early inflammatory response and bursal injury in chicken.

Studies demonstrated that chicken ghrelin mRNA was expressed in immune organs of chicken. However, it was not known for its functions in chicken immune system. This study aimed to investigate the effects of ghrelin on infectious bursal disease virus (IBDV)-induced acute inflammatory and bursal injury. Chickens were divided into 4 groups. One group was used as control ("C"). The other three groups incubated with IBDV on the 19th d, of which 2 were injected intraperitoneally with 0.5 nmol ("LG") or 1.0 nmol ("HG") ghrelin/100g body weight from 18th to 22nd d, respectively, and one was injected intraperitoneally with PBS ("I"). Results showed that cytokines including interleukin (IL)-6, IL-1ß, and IL-8 mRNA expression in I group were upregulated significantly after chickens infected with IBDV from 1 d post-infection (dpi) to 3 dpi (P < 0.05). However, the expression level of IL-6, IL-1ß, and IL-8 mRNA in LG and HG groups was 7.3, ∼43.3% as much as that of the I group at 2 dpi and 3 dpi (P < 0.05). Moreover, ghrelin administration attenuated significantly the bursal injury from 1 dpi to 7 dpi and prevents the reduction of bird weight gain at 5 dpi and 7 dpi, which were induced by IBDV (P < 0.05). The results indicated that ghrelin could play an important role in the immune system of chicken.

Alleviation of infectious-bursal-disease-virus-induced bursal injury by betaine is associated with DNA methylation in IL-6 and interferon regulatory factor 7 promoter.

Infectious bursal disease virus (IBDV) often infects young chickens and causes severe immunosuppression and inflammatory injury. Betaine is an antiviral and anti-inflammatory ingredient that may exert functions through epigenetic regulation. However, the effects of betaine on an IBDV-induced bursal injury and their underlying mechanisms have not been investigated. In this study, betaine was supplemented to the drinking water of newly hatched commercial broilers for 3 wk. Afterward, the chickens were infected with the IBDV. After 5 D of infection, the bursal lesions were examined. The mRNA expression levels of IBDV VP2 gene, pro-inflammatory cytokines, and interferons were detected. Furthermore, the 5-methylcytosine level of the CpG island in the promoter region of IL-6 and interferon regulatory factor 7 (IRF7) were determined. The IBDV induced the depletion of lymphocytes and inflammation in the bursal follicles. IBDV infection considerably elevated the mRNA levels of VP2, IL-6, types I (IFNα and IFNß) and II (IFNγ) interferons, and IRF7. The CpG island methylation in the promoter regions of IL-6 and IRF7 were substantially decreased after IBDV infection. Betaine administration attenuated the IBDV-induced bursal lesions. Meanwhile, the IBDV-induced mRNA expression levels of IL-6, IFNß, and IRF7 were suppressed by betaine consumption. Furthermore, the hypomethylation effects of IBDV infection to the promoter regions of IL-6 and IRF7 genes were eliminated and relieved by betaine administration. Our results indicated that the IBDV-induced expression levels of IL-6 and IRF7 genes are associated with the suppression of methylation in the promoter region. Betaine administration through drinking water may alleviate the IBDV-induced bursal injury via epigenetic regulation.

Intestinal Parasites in the Critically Endangered Przewalski's Gazelle ( Procapra przewalskii ) in China, with the Description of a New Species of Eimeria (Apicomplexa: Eimeriidae).

We used a flotation technique to survey fecal samples from 27 Przewalski's gazelles ( Procapra przewalskii Büchner, 1891) for intestinal parasites. Samples were collected from the Qinghai Lake region, Qinghai-Tibet plateau, China, in April 2015. We report parasites belonging to two nematode (Nematodirus, Marshallagia) genera and one apicomplexan (Eimeria) genus; one of the latter was a new species from the critically endangered Przewalski's gazelle. Sporulated oocysts of the new eimerian were spheroidal; they had a thick, outer wall without a micropyle, but with a distinct polar granule and an oocyst residuum. Oocysts measured a mean of 18.6(±0.8)×17.3(±0.9) µm in length and width, with a mean 1.1(±0.1) length/width ratio and a spheroidal oocyst residuum that had a mean width of 2.0(±0.7) µm. Sporocysts were ovoid, measuring a mean of 9.4(±0.6)×5.5(±0.5) µm in length and width, with a mean 1.7(±0.2) length/width ratio. A Stieda body and sporocyst residuum were present, but a substieda body was absent. Sporozoites each had a small anterior refractile body and a larger, posterior refractile body; a small nucleus was visible between them.

The synergistic effect of density stress during the maternal period and adulthood on immune traits of root vole (Microtus oeconomus) individuals-a field experiment.

The literature reveals that stress in early life or adulthood can influence immune function. As most studies on this are from the laboratory, there is a need for replicated studies in wild animals. This study aims to examine the effects of density stress during the maternal period and adulthood on immune traits of root vole (Microtus oeconomus) individuals. Four replicated high- and low-density parental populations were established, from which we obtained offspring and assigned each into four enclosures, two for each of the two density treatments used in establishing parental populations. The F1 offspring fecal corticosterone metabolite response to acute immobilization stress, anti-keyhole limpet hemocyanin immunoglobulin G (anti-KLH IgG) level, phytohemagglutinin (PHA)-delayed hypersensitivity and hematology at the end of the first breeding season, and prevalence and intensity of coccidial infection throughout the two breeding seasons, were tested. Density-induced maternally stressed offspring had delayed responses to acute immobilization stress. Density-stressed offspring as adults had reduced anti-KLH IgG levels and PHA responses, and the effects further deteriorated in maternally stressed offspring, leading to higher coccidial infection in the first breeding season than in the second. No correlations were found between immune traits or coccidial infection and survival over winter. These findings indicated that the combined density stresses during the maternal period and adulthood exhibited negative synergistic effects on immune traits. The synergistic effects lead to higher coccidial infection; however, this consequently reduced the risk of subsequent infection. The increased coccidial infection mediated by the synergistic effects may have an adaptive value in the context of the environment.

Maternal effects and population regulation: maternal density-induced reproduction suppression impairs offspring capacity in response to immediate environment in root voles Microtus oeconomus.

The hypothesis that maternal effects act as an adaptive bridge in translating maternal environments into offspring phenotypes, and thereby affecting population dynamics has not been studied in the well-controlled fields. In this study, the effects of maternal population density on offspring stress axis, reproduction and population dynamics were studied in root voles (Microtus oeconomus). Parental enclosures for breeding offspring were established by introducing six adults per sex into each of 4 (low density) and 30 adults per sex into each of another 4 (high density) enclosures. Live-trapping started 2 weeks after. Offspring captured at age of 20-30 days were removed to the laboratory, housed under laboratory conditions until puberty, and subsequently used to establish offspring populations in these same enclosures, after parental populations had been removed. [Correction added on 8 January 2015 after first online publication: '10-20 days' has been changed to '20-30 days.'] Offspring from each of the two parental sources were assigned into four enclosures with two for each of the two density treatments used in establishing parental populations (referred to as LL and LH for maternally unstressed offspring, assigned in low and high density, and HL and HH for maternally stressed offspring, assigned in low and high density). Faecal corticosterone metabolites (FCM) levels, offspring reproduction traits and population dynamics were tested following repeated live-trapping over two seasons. Differential fluctuations in population size were observed between maternally density-stressed and density-unstressed offspring. Populations in LL and LH groups changed significantly in responding to initial density and reached the similar levels at beginning of the second trapping season. Populations in HL and HH groups, however, were remained relatively steady, and in HL group, the low population size was sustained until end of experiment. Maternal density stress was associated with FCM elevations, reproduction suppression and body mass decrease at sexual maturity in offspring. The FCM elevations and reproduction suppression were independent of offspring population density and correlated with decreased offspring quality. These findings indicate that intrinsic state alterations induced by maternal stress impair offspring capacity in response to immediate environment, and these alterations are likely mediated by maternal stress system. The maladaptive reproduction suppression seen in HL group suggests intrinsic population density as one of ecological factors generating delayed density-dependent effects.

Four new coccidia (Apicomplexa: Eimeriidae) from the Plateau zokor, Myospalax baileyi Thomas (Rodentia: Myospalacinae), a subterranean rodent from Haibei area, Qinghai Province, China.

Thirty-eight faecal samples from the Plateau zokor, Myospalax baileyi Thomas, collected in the Haibei Area, Qinghai Province, China, were examined for the presence of coccidia (Apicomplexa: Eimeriidae). Seventeen of 38 faecal samples (44.7%) were found to contain coccidian oöcysts representing four new species of Eimeria Schneider, 1875, and four of 17 (23.5%) infected zokors were concurrently infected with two or three of these eimerian species. The sporulated oöcysts of Eimeria myospalacensis n. sp. are ovoidal, 9.5-17.0 × 8.0-13.0 (mean 13.0 × 10.4) µm; a polar granule is present, oöcyst residuum is absent; sporocysts are ovoidal, 4.5-7.5 × 3.0-5.0 (mean 6.3 × 4.2) µm and have both a Stieda body and residuum. Oöcysts of Eimeria fani n. sp. are ellipsoidal to cylindroidal, 12.5-16.0 × 8.0-11.0 (mean 14.6 × 9.9) µm; a polar granule is present, but micropyle and residuum are lacking; sporocysts are ovoidal, 4.5-7.5 × 3.0-5.3 (mean 6.7 × 4.4) µm; a residuum and a Steida body are present. Oöcysts of Eimeria baileyii n. sp. are ellipsoidal, 15.0-23.0 × 12.0-18.0 (mean 18.2 × 13.7) µm; a polar granule is present but oöcyst residuum is absent; sporocysts are ovoidal, 8.0-11.0 × 5.0-7.0 (mean 9.5 × 5.9) µm and have both a Stieda body and residuum. Oöcysts of Eimeria menyuanensis n. sp. are ovoidal, 12.5-21.0 × 11.0-18.0 (mean 17.1 × 14.6) µm, with a distinct micropyle c.2.5 µm wide; a polar granule is present but a residuum is absent; sporocysts are ovoidal, 8.0-12.0 × 5.0-7.0 (mean 10.2 × 6.4) µm, and have both a Stieda body and residuum.

Chronic hypoxia exposure during pregnancy is associated with a decreased active nursing activity in mother and an abnormal birth weight and postnatal growth in offspring of rats.

Stress during pregnancy is known to have a significant impact on animal's behavior and offspring development. The effects of gestational hypoxia on maternal behavior have not been studied. In the present study, we investigated the effects of gestational hypoxia exposure on dam's maternal behavior, offspring's growth and plasma corticosterone levels after parturition in rats. Altitude hypoxia (3 and 5 km) was simulated in the hypobaric chambers during the last week of pregnancy and the effects were compared to those found in controls exposed at sea level. We found that gestational hypoxia significantly decreased dam's arched-back nursing activity across the lactation period. The effect was more profound in 5 km group. Gestational hypoxia also altered other maternal behaviors such as blanket and passive nursing. Hypoxia exposure was associated with abnormal birth weight and postnatal growth in pups, with a significantly higher and lower birth weight than control found in 3 and 5 km groups, respectively, and accelerated growth in both stressed groups. Gestational hypoxia exposure significantly elevated plasma corticosterone levels in dams at the time of weaning and in pups across the measurement days. Taken together, the present results indicate that hypoxia, particularly severe hypoxia during the late phase of pregnancy has a significantly adverse impact on animal's behavior, endocrine function and offspring development. The higher birth weight found in the offspring of 3 km group suggests a compensatory system counteracting with the inhibitory effects of hypoxia on fetus growth at this altitude.