Meteorological, demographic, and environmental risk factors for Leishmania infantum in wildlife.

Leishmania infantum is an important and neglected vector-borne zoonotic protozoa endemic in the Mediterranean basin. Several wild and domestic mammals can contribute to maintaining its circulation but their importance as effective reservoirs is still under discussion and varies depending on local ecological communities. By combining environmental, climatic, and individual information, this study assessed the presence of L. infantum DNA in a set of wild species from Northwestern Italy and the potential ecological factors related to the risk of infection. From 2020 to 2022, 304 free-ranging wild animals were analyzed for the detection of L. infantum DNA in the spleen and popliteal lymph node (when available). The prevalence obtained in wild boar (Sus scrofa) and roe deer (Capreolus capreolus) was higher than those previously reported (% ± confidence interval 95%; 42.9 ± 18.4% and 27 ± 6.6% in wild boar and roe deer, respectively), and this is the first report of this parasite infecting the coypu Myocastor coypus (60 ± 34.7%). L. infantum DNA was detected in all the seasons including those free of adult sandflies and seasonal differences were minimal, suggesting a long course of infection. The models revealed that animals from rainy areas with higher greenness during the summer, highly populated by humans and predominantly covered by water surfaces had a higher risk of L. infantum. This study contributes to confirming previous findings on the existence of a sylvatic cycle for L. infantum in certain regions of Italy, as well as on the potential epidemiological role of roe deer for this parasite given the elevated prevalence found.

Camera traps reveal a high prevalence of sarcoptic mange in red foxes from northern Spain.

The mite Sarcoptes scabiei affects numerous mammal species causing the sarcoptic mange, a widespread disease with relevance for wildlife conservation, welfare, and management. The red fox (Vulpes vulpes) could become infested by direct and indirect routes leading to external skin lesions potentially recognizable by devices such as camera traps (CTs). In the present study, 86 randomly placed CTs were used to investigate the apparent prevalence and severity of S. scabiei in a red fox population from northern Spain. Their potential environmental and population-related drivers were also assessed. A total of 341 independent encounters were examined to visually identify mange-compatible lesions. The apparent prevalence was 19.16% (confidence interval (CI) 95%: 15.08-23.80) of which 82.81% (CI95%: 71.33-91.10) were severe. Our results revealed that habitat attributes such as lower altitudes, higher coverage of water-linked habitats and woodland predominance, were significant predictors of the apparent risk of mange. The models also suggested that the apparent prevalence of mange was associated with poor body condition and elevated frequencies of spatial coincidence among fox encounters, which facilitates indirect transmission. Interestingly, we did not observe mange-compatible lesions in other sympatric wild species (>15,000 encounters examined). This could be explained by the mite's host specificity and the low probability that these other potential hosts use sites where transmission among foxes usually occurs, such as dens. This study illustrates how camera trapping can be used as an interesting tool for the surveillance of wildlife diseases, thus overcoming the logistic constraints derived from direct sampling and allowing the early detection and better management of pathogens in the riskiest areas.

Extract of Cimicifuga racemosa (L.) Nutt protects ovarian follicle reserve of mice against in vitro deleterious effects of dexamethasone.

The present study aims to investigate if Cimicifuga racemosa (L.) Nutt extract (CIMI) reduces deleterious effects of dexamethasone (DEXA) in ovaries cultured in vitro. Mouse ovaries were collected and cultured in DMEM+ only or supplemented with 5 ng/mL of CIMI, or 4 ng/mL DEXA, or both CIMI and DEXA. The ovaries were cultured at 37.5°C in 5% CO2 for 6 days. Ovarian morphology, follicular ultrastructure, and the levels of mRNA for Bax, Bcl-2, and Caspase-3 were evaluated. The results showed that DEXA reduced the percentage of morphologically normal follicles, while CIMI prevented the deleterious effects caused by DEXA. In addition, DEXA negatively affected the stromal cellular density, while CIMI prevented these adverse effects. Ovaries cultured with DEXA and CIMI showed similar levels of mRNA for Bax, Bcl-2, and Caspase-3 compared to those cultured in control medium, while ovaries cultured with DEXA had increased expression of the above genes. Additionally, the ultrastructure of the ovaries cultured with CIMI was well preserved. Thus, the extract of CIMI was able to prevent the deleterious effects caused by DEXA on cultured mouse ovaries.

Using integrated wildlife monitoring to prevent future pandemics through one health approach.

In the One Health context, Integrated Wildlife Monitoring (IWM) merges wildlife health monitoring (WHM) and host community monitoring to early detect emerging infections, record changes in disease dynamics, and assess the impact of interventions in complex multi-host and multi-pathogen networks. This study reports the deployment and results obtained from a nationwide IWM pilot test in eleven sites representing the habitat diversity of mainland Spain. In each study site, camera-trap networks and sampling of indicator species for antibody and biomarker analysis were used to generate information. The results allowed identifying differences in biodiversity and host community characteristics among the study sites, with a range of 8 to 19 relevant host species per point. The Eurasian wild boar (Sus scrofa) was the most connected and central species of the host communities, becoming a key target indicator species for IWM. A negative relationship between biodiversity and disease risk was detected, with a lower number and prevalence of circulating pathogens in the sites with more species in the community and larger network size. However, this overall trend was modified by specific host-community and environmental factors, such as the relative index of wild boar - red deer interactions or the proximity to urban habitats, suggesting that human-driven imbalances may favour pathogen circulation. The effort of incorporating wildlife population monitoring into the currently applied WHM programs to achieve effective IWM was also evaluated, allowing to identify population monitoring as the most time-consuming component, which should be improved in the future. This first nationwide application of IWM allowed to detect drivers and hotspots for disease transmission risk among wildlife, domestic animals, and humans, as well as identifying key target indicator species for monitoring. Moreover, anthropogenic effects such as artificially high wildlife densities and urbanisation were identified as risk factors for disease prevalence and interspecific transmission.

Extract of Cimicifuga racemosa (L.) Nutt protects ovarian follicle reserve of mice against in vitro deleterious effects of dexamethasone

The present study aims to investigate if Cimicifuga racemosa (L.) Nutt extract (CIMI) reduces deleterious effects of dexamethasone (DEXA) in ovaries cultured in vitro. Mouse ovaries were collected and cultured in DMEM+ only or supplemented with 5 ng/mL of CIMI, or 4 ng/mL DEXA, or both CIMI and DEXA. The ovaries were cultured at 37.5°C in 5% CO2 for 6 days. Ovarian morphology, follicular ultrastructure, and the levels of mRNA for Bax, Bcl-2, and Caspase-3 were evaluated. The results showed that DEXA reduced the percentage of morphologically normal follicles, while CIMI prevented the deleterious effects caused by DEXA. In addition, DEXA negatively affected the stromal cellular density, while CIMI prevented these adverse effects. Ovaries cultured with DEXA and CIMI showed similar levels of mRNA for Bax, Bcl-2, and Caspase-3 compared to those cultured in control medium, while ovaries cultured with DEXA had increased expression of the above genes. Additionally, the ultrastructure of the ovaries cultured with CIMI was well preserved. Thus, the extract of CIMI was able to prevent the deleterious effects caused by DEXA on cultured mouse ovaries.

Immunolocalization of melatonin receptors in bovine ovarian follicles and in vitro effects of melatonin on growth, viability and gene expression in secondary follicles.

This study aims to investigate the (1) expression of melatonin receptors types 1A/B (MTNR1A/B) in bovine ovaries and (2) the in vitro effects of melatonin on secondary follicle development, antrum formation, viability, and expression of messenger ribonucleic acid (mRNA) for superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase-1 (GPX1) and peroxiredoxin 6 (PRDX6). The expression of MTNR1A/B in bovine ovarian follicles was demonstrated by immunohistochemistry. To choose the most effective concentration of melatonin on follicular growth and viability, isolated secondary follicles were cultured individually at 38.5°C, with 5% CO2 in air, for 18 d in TCM-199+ alone or supplemented with 10-11, 10-9, 10-7 or 10-5 M melatonin. Then, melatonin receptor antagonist, luzindole, was tested to further evaluate the mechanisms of actions of melatonin, that is, the follicles were cultured in control medium alone or supplemented with 10-7 M melatonin, 10 µM luzindole and both 10-7 M melatonin and 10 µM luzindole. Follicular growth, morphology and antrum formation were evaluated at days 6, 12 and 18. At the end of culture, viability of secondary follicles was analyzed by calcein-AM and ethidium homodimer-1, and the relative levels of mRNA for SOD, CAT, GPX1 and PRDX6 were evaluated by real time polymerase chain reaction. Immunohistochemistry results showed expression of MTNR1A/B in oocyte and granulosa cells of primordial, primary, secondary and antral follicles. Secondary follicles cultured in medium supplemented with melatonin at different concentrations had well preserved follicles after 18 d of culture. Furthermore, follicles cultured in presence of 10-7 M melatonin presented significantly higher diameters than those cultured in other treatments. The presence of melatonin receptor antagonist, luzindole, blocked the effects of melatonin on follicular growth and viability. In addition, follicles cultured in medium containing only melatonin had significantly higher rates of antrum formation. Follicles cultured in medium containing only melatonin had higher relative levels of mRNA for CAT, SOD and PRDX-6 than those cultured with both melatonin and luzindole. Follicles cultured with luzindole only or both melatonin and luzindole had lower relative levels of mRNA for PRDX6 and GPX1 than those cultured control medium. In conclusion, melatonin promotes growth of bovine secondary follicles through its membrane-coupled receptors, while luzindole blocks the effects of melatonin on follicle growth and reduces the expression of antioxidant enzymes in cultured follicles.

Number needed to screen for TB in clinical, structural or occupational risk groups.

BACKGROUND: Screening for active TB using active case-finding (ACF) may reduce TB incidence, prevalence, and mortality; however, yield of ACF interventions varies substantially across populations. We systematically reviewed studies reporting on ACF to calculate the number needed to screen (NNS) for groups at high risk for TB.METHODS: We conducted a literature search for studies reporting ACF for adults published between November 2010 and February 2020. We determined active TB prevalence detected through various screening strategies and calculated crude NNS for - TB confirmed using culture or Xpert® MTB/RIF, and weighted mean NNS stratified by screening strategy, risk group, and country-level TB incidence.RESULTS: We screened 27,223 abstracts; 90 studies were included (41 in low/moderate and 49 in medium/high TB incidence settings). High-risk groups included inpatients, outpatients, people living with diabetes (PLWD), migrants, prison inmates, persons experiencing homelessness (PEH), healthcare workers, and miners. Screening strategies included symptom-based screening, chest X-ray and Xpert testing. NNS varied widely across and within incidence settings based on risk groups and screening methods. Screening tools with higher sensitivity (e.g., Xpert, CXR) were associated with lower NNS estimates.CONCLUSIONS: NNS for ACF strategies varies substantially between adult risk groups. Specific interventions should be tailored based on local epidemiology and costs.

A systematic review of the number needed to screen for active TB among people living with HIV.

BACKGROUND: Systematic screening for active TB is recommended for all people living with HIV (PLWH); however, case detection remains poor globally. We investigated the yield of active case finding (ACF) by calculating the number needed to screen (NNS) to detect a case of active TB among PLWH.METHODS: We identified studies reporting ACF for TB among PLWH published from November 2010 to February 2020. We calculated crude NNS for Xpert- or culture-confirmed TB and weighted mean NNS stratified by screening approach, population/risk group, and country TB burden.RESULTS: Of the 27,221 abstracts screened, we identified 58 studies eligible for inclusion, including 5 in low/moderate TB incidence settings and 53 in medium/high incidence settings. Populations screened for TB included inpatients, outpatients not receiving antiretroviral therapy (ART), outpatients receiving ART, those with CD4 < 200 cells/µL, children aged ≤15 years, pregnant PLWH, and PLWH in prisons. Screening tools included symptom-based screening, chest X-ray, C-reactive protein levels, and Xpert. The weighted mean NNS varied across groups but was consistently low, ranging from 4 among inpatients in moderate/high TB burden settings to 137 among pregnant PLWH in moderate/high TB burden settings.CONCLUSIONS: ACF is a high yield intervention among PLWH. Approaches to screening should be tailored to local epidemiological and health-system contexts, and sensitive screening tools such as Xpert should be implemented where feasible.

Eugenol influences the expression of messenger RNAs for superoxide dismutase and glutathione peroxidase 1 in bovine secondary follicles cultured in vitro.

This study aimed to investigate the effects of eugenol on growth, viability, antrum formation and mRNA expression of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase 1 (GPX1) and peroxiredoxin 6 (PRDX6) in bovine secondary follicles cultured in vitro. To this end, bovine ovaries were collected from a local slaughterhouse and in the laboratory the follicles were isolated from the ovarian cortex. The follicles were then cultured in TCM-199+ alone or supplemented with different concentrations of eugenol (0.5, 5.0 and 50.0 µM). Follicular diameters and antrum formation were evaluated on days 0, 6, 12 and 18. Viability analysis was performed using calcein and ethidium homodimer. Real-time PCR was used to quantify mRNA levels for SOD, CAT, GPX1 and PRDX6 in cultured follicles. Follicular diameters and mRNA levels in follicles cultured in vitro were compared using analysis of variance and Kruskal-Wallis tests, while follicular survival and antrum formation were compared using the chi-squared test (P < 0.05). The results showed that secondary follicles cultured with eugenol maintained similar morphology and viability to follicles cultured in the control group. A progressive increase in follicular diameter was observed between days 0 and 12 for all treatments, except for follicles cultured with 50 µM eugenol. Eugenol (5.0 and 50.0 µM) increased mRNA levels for GPX1 in cultured follicles, but 0.5 µM eugenol reduced mRNA levels for SOD. The addition of eugenol did not influence mRNA expression for CAT and PRDX6. In conclusion, eugenol supplementation reduces mRNA levels for SOD and increases mRNA levels of GPX1 in bovine secondary follicles cultured in vitro.

Effects of dexamethasone on growth, viability and ultrastructure of bovine secondary follicles cultured in vitro.

This study aimed to evaluate the effects of dexamethasone on development, viability, antrum formation and ultrastructural integrity of bovine secondary follicles cultured in vitro for 18 days. Bovine ovaries were obtained from slaughterhouses and secondary follicles of ~150-200 µm diameter were isolated and cultured in the laboratory in TCM-199+ alone or supplemented with different concentrations of dexamethasone (1, 10, 100 and 1000 ng/ml). Follicle viability was evaluated after the culture period, using calcein-AM (viable) and ethidium homodimer (nonviable). Follicle diameters and antrum formation were evaluated at days 0, 6, 12 and 18. Before or after in vitro culture, follicles were fixed for histological and ultrastructural analysis. Follicle diameters were evaluated using analysis of variance and Kruskal-Wallis test, while chi-squared test was used to evaluate the percentage of viable follicles and antrum formation (P < 0.05). Follicles cultured for 6 days with all treatments increased their diameters significantly, but there was no significant difference between treatments at the end of the culture period. In vitro cultured follicles showed antral cavity formation at the end of the culture period, but no influence of dexamethasone was seen. Ultrastructural analysis showed that follicles cultured with dexamethasone (1, 10, 100 and 1000 ng/ml) had well preserved granulosa cells. However, oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone showed signs of degeneration. It can be concluded that follicles cultured in vitro in the presence of dexamethasone demonstrated continuous in vitro growth, but oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone had poor ultrastructure.

Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources during in vitro culture.

The present study evaluated the effect of knockout serum replacement (KSR), fetal bovine serum (FBS) and bovine serum albumin (BSA) on the viability and growth of bovine secondary follicles cultured in vitro for 12 days. To this end, secondary follicles were isolated (185-202 µm) and cultured in vitro in TCM-199+ medium supplemented with KSR (5% and 10%), FBS (5% and 10%) or BSA (3 mg/ml) at 38.5°C with 5% CO2 in air. Follicular diameters were evaluated on days 0, 4, 8 and 12. After 12 days of culture, follicular survival analysis was performing by using calcein-AM and ethidium homodimer. Before and after culture, follicles were fixed in paraformaldehyde for histological evaluation. Follicular diameter at different days of culture were compared using the Kruskal-Wallis test, while the percentages of viable follicles were analyzed by chi-squared test (P < 0.05). Results showed that follicles cultured in the presence of KSR at both concentrations presented higher follicular survival rates than those cultured in control medium alone or supplemented with FBS or BSA. Conversely, the presence of KSR, BSA or FBS did not increase follicular diameter after 12 days of culture. Histology analysis showed that, among the tested treatments, follicles cultured in the presence of KSR had preserved rounded oocytes, juxtaposed granulosa cells and intact basal membrane. In conclusion, supplementation of culture medium with KSR increases the follicular survival of bovine secondary follicles cultured in vitro.

Effects of epidermal growth factor and progesterone on development, ultrastructure and gene expression of bovine secondary follicles cultured in vitro.

The aims of this study were to investigate the effects of epidermal growth factor (EGF) and progesterone on the development, viability and the gene expression of bovine secondary follicle culture in vitro for 18 days. Secondary follicles (∼0.2 mm) were isolated from ovarian cortex and individually cultured at 38.5 °C, with 5% CO2 in air, for 18 days, in TCM-199+ (n = 63) alone (control medium) or supplemented with 10 ng/mL progesterone (n = 64), 10 ng/mL EGF (n = 61) or both EGF and progesterone (n = 66). The effects of these treatments on growth, antrum formation, viability, ultrastructure and mRNA levels for GDF-9, c-MOS, H1foo and cyclin B1 were evaluated, significantly different (p < 0.05). The results showed that there was a progressive increase in follicular diameter in all treatments, but only follicles cultured in medium supplemented with EGF had increased significantly in diameter when compared to follicles cultured in the control medium at the end of the culture period, significantly different (p < 0.05). A positive interaction between EGF and progesterone was not observed. In addition, the presence of EGF, progesterone or both in culture medium did not influence the rate of follicle survival and antrum formation. However, the presence of only progesterone in cultured medium increased the expression of mRNAs for GDF9 and cyclin B1 in oocytes. EGF also significantly increased the levels of mRNAs for cMOS and GDF9 when compared to follicles cultured in control medium. Ultrastructural analyzes showed that cultured follicles in all treatments maintained the integrity of granulosa cells. In conclusion, the EGF promotes the development of secondary follicles cultured in vitro for 18 days and increases the expression of cMOS and GDF9, while progesterone alone or in association with EGF have not a positive effect on follicular growth. However, progesterone increases the expression of GDF9 and cyclin B1 in oocytes.

Laser-driven shock compression of "synthetic planetary mixtures" of water, ethanol, and ammonia.

Water, methane, and ammonia are commonly considered to be the key components of the interiors of Uranus and Neptune. Modelling the planets' internal structure, evolution, and dynamo heavily relies on the properties of the complex mixtures with uncertain exact composition in their deep interiors. Therefore, characterising icy mixtures with varying composition at planetary conditions of several hundred gigapascal and a few thousand Kelvin is crucial to improve our understanding of the ice giants. In this work, pure water, a water-ethanol mixture, and a water-ethanol-ammonia "synthetic planetary mixture" (SPM) have been compressed through laser-driven decaying shocks along their principal Hugoniot curves up to 270, 280, and 260 GPa, respectively. Measured temperatures spanned from 4000 to 25000 K, just above the coldest predicted adiabatic Uranus and Neptune profiles (3000-4000 K) but more similar to those predicted by more recent models including a thermal boundary layer (7000-14000 K). The experiments were performed at the GEKKO XII and LULI2000 laser facilities using standard optical diagnostics (Doppler velocimetry and optical pyrometry) to measure the thermodynamic state and the shock-front reflectivity at two different wavelengths. The results show that water and the mixtures undergo a similar compression path under single shock loading in agreement with Density Functional Theory Molecular Dynamics (DFT-MD) calculations using the Linear Mixing Approximation (LMA). On the contrary, their shock-front reflectivities behave differently by what concerns both the onset pressures and the saturation values, with possible impact on planetary dynamos.

High prevalence and intensity of Stephanurus dentatus in a population of wild boar (Sus scrofa) in south western Spain.

In the period from October 2016 to February 2017, the urinary tracts of 390 wild boar (Sus scrofa) from four areas of south central Spain (102 from Doñana National Park; 150 from Sierra Morena and the Toledo Mountains; 84 from Sierra Nevada; 54 from Sierra de Cazorla, Segura y Las Villas Natural Park) were examined for the presence of adult specimens of Stephanurus dentatus (Nematoda: Strongyloidea). This parasite was only detected in the wild boar population of Doñana National Park, with high prevalence (76.5±4.2%; 78/102), mean intensity (43.2±4.4) and mean abundance (33.1±3.8). Juvenile wild boar had significantly lower prevalence and abundance than subadult and adult wild boar. The intensity of infestation was significantly higher in male than in female wild boar. The detection of a focus of S. dentatus infestation in the wild boar population in Doñana National Park will provide further opportunities for understanding the epidemiology of this parasite.

Spontaneous fertility in a male patient with testotoxicosis despite suppression of FSH levels.

Testotoxicosis is a rare cause of peripheral precocious puberty in boys caused by constitutively activating mutations of the LHCG receptor. Affected males usually have normal gonadotropin profiles and fertility in their adult life. Here, we described the long-term follow-up of a 24-year-old young man with severe testotoxicosis due to a de novo activating mutation in the third transmembrane helix of the LHCGR (p.Leu457Arg). This patient was treated with different medications, including medroxyprogesterone acetate, ketoconazole, cyproterone acetate and aromatase inhibitor from age 2.5 to 9.5 years. His basal and GnRH-stimulated gonadotropin levels were continually suppressed during and after medical treatment. At adulthood, extremely high serum testosterone levels (>35 nmol/L), undetectable gonadotropin levels (LH < 0.15 IU/L and FSH < 0.6 IU/L) and oligozoospermia were evidenced. Despite his suppressed FSH levels and an unfavorable spermogram, the patient fathered a healthy girl and biological paternity was confirmed through analysis of microsatellites. Spontaneous fertility in a young man with severe testotoxicosis and chronic suppression of FSH levels reinforces the key role of high intratesticular testosterone levels in human spermatogenesis.

Genetic effects of in vitro germination and plantlet development in chilli pepper.

The objective of this study was to estimate the genetic effects on in vitro germination and development of chili peppers. For this, four genotypes (UFPB-132, -134, -137, and -390) were used as parents. They were crossed in a complete diallel scheme. The parents (4) and hybrid (12) seeds were germinated in glass bottles containing MS media. The following variables were evaluated: seed germination percentage, deformed seedlings percentage, radicle emission percentage, root length, root number, hypocotyl length, hypocotyl width, definitive leaf number, leaf length, and leaf width. The data were submitted to analysis of variance and the means were grouped by the Scott-Knott criteria (P ≤ 0.05). The diallel analysis was performed according to the Griffing method, method I, fixed model. Significant differences for all analysed variables were observed. Additive and non-additive effects were observed influencing the performance of the genotypes in relation to the evaluated variables. The genitor 132 showed the highest general combining ability for almost all evaluated characteristics, except seed germination percentage and deformed seedlings percentage. This study revealed dominance effects are responsible for genotypic variation for almost all evaluated traits. In addition, we found significant reciprocal effects for all studied characters. For the traits to which non-additive variances were important (germination percentage, deformed seedlings percentage, root number, hypocotyl length, hypocotyl width, and definitive leaf number, leaf length, and leaf width), there was an additional opportunity for developing F1 hybrid.

Genetic diversity of arboreal cotton populations of the Brazilian semiarid: a remnant primary gene pool for cotton cultivars.

Mocó cotton belong to the same species as the cultivated species, Gossypium hirsutum, and cultivated forms were mainly landraces but also developed as cultivars, bearing good fiber quality and drought tolerant when cropped as a perennial species. The northeast Brazil crop system based on this cotton type is finished, with a few small area planted in the three main States, where it was previously cultivated (Ceará, Paraíba, and Rio Grande do Norte), but in others, maintenance is accomplished by single dooryard plants. Plants were found in all visited Northeast Brazil municipalities, sometimes in the North of the country, and were collected for ex situ preservation and evaluation. Most of seeds had no fuzz (62.2%) and 94.6% of the genotypes presented spot in flowers. Seventy-one alleles were revealed in 12 loci. The genetic structure of the population evaluated by microsatellite markers shows two main groups, one comprising the Seridó region where landraces were originated and other comprising the state of Ceará, where a specific breeding program was developed. Genotypes collected in North Brazil States as well as those collected in Bahia, Alagoas, and Sergipe grouped with those collected in Ceará. The Mantel correlogram indicates a significant (P < 0.05) correlation between genetic and geographical distances up to 77 km. The ex situ maintenance and agronomical evaluation are the main concerns for mocó, as the use of the agricultural interesting traits, possibly introgressed to other genotypes, is predicted. The in situ preservation is still of interest since there is more diversity there than in the collected plants and some should be continued due to use as medicinal plant.

Characterization and genetic diversity of pepper (Capsicum spp) parents and interspecific hybrids.

Pepper species exhibit broad genetic diversity, which enables their use in breeding programs. The objective of this study was to characterize the diversity between the parents of different species and their interspecific hybrids using morphological and molecular markers. The parents of Capsicum annuum (UFPB-01 and -137), C. baccatum (UFPB-72), and C. chinense (UFPB-128) and their interspecific hybrids (01x128, 72x128, and 137x128) were used for morphological and molecular characterization. Fruit length and seed yield per fruit (SYF) traits showed the highest variability, and three groups were formed based on these data. CVg/CVe ratio values (>1.0) were calculated for leaf length (1.67) and SYF (5.34). The trait that most contributed to divergence was the largest fruit diameter (26.42%), and the trait that least contributed was pericarp thickness (0.33%), which was subject to being discarded. The 17 primers produced 58 polymorphic bands that enabled the estimation of genetic diversity between parents and hybrids, and these results confirmed the results of the morphological data analyses. The principal component analysis results also corroborated the morphological and random-amplified polymorphic DNA data, and three groups that contained the same individuals were identified. These results confirmed reports in the literature regarding the phylogenetic relationships of the species used as parents, which demonstrated that C. annuum was closer to C. chinense as compared to C. baccatum.

The TcTASV proteins are novel promising antigens to detect active Trypanosoma cruzi infection in dogs.

In regions where Chagas disease is endemic, canine Trypanosoma cruzi infection is highly correlated with the risk of transmission of the parasite to humans. Herein we evaluated the novel TcTASV protein family (subfamilies A, B, C), differentially expressed in bloodstream trypomastigotes, for the detection of naturally infected dogs. A gene of each TcTASV subfamily was cloned and expressed. Indirect enzyme-linked immunosorbent assays (ELISA) were developed using recombinant antigens individually or mixed together. Our results showed that dogs with active T. cruzi infection differentially reacted against the TcTASV-C subfamily. The use of both TcTASV-C plus TcTASV-A proteins (Mix A+C-ELISA) enhanced the reactivity of sera from dogs with active infection, detecting 94% of the evaluated samples. These findings agree with our previous observations, where the infected animals exhibited a quick anti-TcTASV-C antibody response, coincident with the beginning of parasitaemia, in a murine model of the disease. Results obtained in the present work prove that the Mix A+C-ELISA is a specific, simple and cheap technique to be applied in endemic areas in screening studies. The Mix A+C-ELISA could help to differentially detect canine hosts with active infection and therefore with high impact in the risk of transmission to humans.

Protein and messenger RNA expression of interleukin 1 system members in bovine ovarian follicles and effects of interleukin 1ß on primordial follicle activation and survival in vitro.

This study aimed to investigate the expression of interleukin 1 (IL-1) system members (proteins and messenger RNA of ligands and receptors) and its distribution in ovarian follicles of cyclic cows and to evaluate the effects of IL-1ß on the survival and activation of primordial follicles in vitro. The ovaries were processed for localization of IL-1 system in preantral and antral follicles by immunohistochemical, real-time polymerase chain reaction, and Western blot analysis. For in vitro studies, ovarian fragments were cultured in α-MEM(+) supplemented with IL-1ß (0, 1, 10, 50, or 100 ng/mL), and after 6 d, the cultured tissues were processed for histologic analysis. Immunohistochemical results showed that the IL-1 system proteins IL-1ß, IL-1RA, IL-1RI, and IL-1RII were detected in the cytoplasm of oocytes and granulosa cells from all follicular categories and theca cells of antral follicles. Variable levels of messenger RNA for the IL-1 system members were observed at different stages of development. After 6 d of culture, the presence of IL-1ß (10 or 50 ng/mL) was effective in maintaining the percentage of normal follicles and in promoting primordial follicle activation. In conclusion, IL-1 system members are differentially expressed in ovarian follicles according to their stage of development. Moreover, IL-1ß promotes the development of primordial follicles. These results indicate an important role of the IL-1 system in the regulation of bovine folliculogenesis.