Eosinophils mediate SIgA production triggered by TLR2 and TLR4 to control Ascaris suum infection in mice.

Human ascariasis is the most prevalent but neglected tropical disease in the world, affecting approximately 450 million people. The initial phase of Ascaris infection is marked by larval migration from the host's organs, causing mechanical injuries followed by an intense local inflammatory response, which is characterized mainly by neutrophil and eosinophil infiltration, especially in the lungs. During the pulmonary phase, the lesions induced by larval migration and excessive immune responses contribute to tissue remodeling marked by fibrosis and lung dysfunction. In this study, we investigated the relationship between SIgA levels and eosinophils. We found that TLR2 and TLR4 signaling induces eosinophils and promotes SIgA production during Ascaris suum infection. Therefore, control of parasite burden during the pulmonary phase of ascariasis involves eosinophil influx and subsequent promotion of SIgA levels. In addition, we also demonstrate that eosinophils also participate in the process of tissue remodeling after lung injury caused by larval migration, contributing to pulmonary fibrosis and dysfunction in re-infected mice. In conclusion, we postulate that eosinophils play a central role in mediating host innate and humoral immune responses by controlling parasite burden, tissue inflammation, and remodeling during Ascaris suum infection. Furthermore, we suggest that the use of probiotics can induce eosinophilia and SIgA production and contribute to controlling parasite burden and morbidity of helminthic diseases with pulmonary cycles.

Genetic Background Affects the Mucosal Secretory IgA Levels, Parasite Burden, Lung Inflammation, and Mouse Susceptibility to Ascaris suum Infection.

Ascariasis is a neglected tropical disease that is widespread in the world and has important socioeconomic impacts. The presence of various stages of worm development in the pulmonary and intestinal mucosae induces a humoral and cellular immune response. However, although there is much evidence of the protective role of mucosal immunity against various pathogens, including helminths, there is still a gap in the knowledge about the immune response and the mechanisms of action that are involved in protection against diseases, especially in the initial phase of ascariasis. Thus, the aim of this study was to evaluate the kinetic aspects of the immune parasitological parameters in intestinal and pulmonary mucosae in male mice with early ascariasis. Therefore, two mouse strains that showed different susceptibilities to ascariasis (BALB/c and C57BL/6J) when experimentally infected with 2,500 infective eggs of Ascaris suum from time point 0 were examined: the immune parasitological parameters were evaluated each 2 days after infection over a period of 12 days. The results were suggestive of a synergetic action of intestinal and pulmonary secretory IgA (S-IgA) contributing to protection against early ascariasis by reducing the amount of migrating larvae as well as the influx of leukocytes in the lung and the consequent impairment of pulmonary capacity.

Nitric oxide contributes to liver inflammation and parasitic burden control in Ascaris suum infection.

BACKGROUND: Human ascariasis is one of the most prevalent neglected tropical diseases worldwide. The immune response during human ascariasis is characterized by Th2 polarization and a mixed Th2/Th17 response during the pathogenesis of experimental larval ascariasis. Cytokines and other pro-inflammatory mediators, such as nitric oxide (NO), are involved in helminthic infections. However, the role of NO in ascariasis remains unclear. OBJECTIVES: Given the importance of NO in inflammation, we aimed to determine the immunological and histopathological alterations in the livers of C57BL/6 iNOS-/- mice during A. suum infection. METHODS: In this study, parasitic load was evaluated in the livers of wild type C57BL/6 and C57BL/6 iNOS-/- mice infected with A. suum. Histopathological and morphometric analyses and analysis of serum cytokines via Cytometric Bead Array were performed, and the activity of eosinophil peroxidase and myeloperoxidase of neutrophils in the tissues were determined. RESULTS: The results showed that NO is important for controlling parasitic load during infection by A. suum. C57BL/6iNOS-/- mice showed reduced inflammatory processes and less tissue damage during liver larval migration of A. suum, which is associated with a reduction in serum levels of pro-inflammatory cytokines. CONCLUSIONS: We demonstrated that NO is a crucial inflammatory molecule during Ascaris sp. infection and controls the establishment of the parasite and the development of the host immune response in the liver.

Concomitant experimental coinfection by Plasmodium berghei NK65-NY and Ascaris suum downregulates the Ascaris-specific immune response and potentiates Ascaris-associated lung pathology.

BACKGROUND: Ascariasis and malaria are highly prevalent parasitic diseases in tropical regions and often have overlapping endemic areas, contributing to high morbidity and mortality rates in areas with poor sanitary conditions. Several studies have previously aimed to correlate the effects of Ascaris-Plasmodium coinfections but have obtained contradictory and inconclusive results. Therefore, the present study aimed to investigate parasitological and immunopathological aspects of the lung during murine experimental concomitant coinfection by Plasmodium berghei and Ascaris suum during larvae ascariasis. METHODS: C57BL/6J mice were inoculated with 1 × 104 P. berghei strain NK65-NY-infected red blood cells (iRBCs) intraperitoneally and/or 2500 embryonated eggs of A. suum by oral gavage. P. berghei parasitaemia, morbidity and the survival rate were assessed. On the seventh day postinfection (dpi), A. suum lung burden analysis; bronchoalveolar lavage (BAL); histopathology; NAG, MPO and EPO activity measurements; haematological analysis; and respiratory mechanics analysis were performed. The concentrations of interleukin (IL)-1ß, IL-12/IL-23p40, IL-6, IL-4, IL-33, IL-13, IL-5, IL-10, IL-17A, IFN-γ, TNF and TGF-ß were assayed by sandwich ELISA. RESULTS: Animals coinfected with P. berghei and A. suum show decreased production of type 1, 2, and 17 and regulatory cytokines; low leukocyte recruitment in the tissue; increased cellularity in the circulation; and low levels of NAG, MPO and EPO activity that lead to an increase in larvae migration, as shown by the decrease in larvae recovered in the lung parenchyma and increase in larvae recovered in the airway. This situation leads to severe airway haemorrhage and, consequently, an impairment respiratory function that leads to high morbidity and early mortality. CONCLUSIONS: This study demonstrates that the Ascaris-Plasmodium interaction is harmful to the host and suggests that this coinfection may potentiate Ascaris-associated pathology by dampening the Ascaris-specific immune response, resulting in the early death of affected animals.

Immunological underpinnings of Ascaris infection, reinfection and co-infection and their associated co-morbidities.

Human ascariasis is the most common and prevalent neglected tropical disease and is estimated that ~819 million people are infected around the globe, accounting for 0.861 million years of disability-adjusted life years in 2017. Even with the existence of highly effective drugs, the constant presence of infective parasite eggs in the environment contribute to a high reinfection rate after treatment. Due to its high prevalence and broad geographic distribution Ascaris infection is associated with a variety of co-morbidities and co-infections. Here, we provide data from both experimental models and humans studies that illustrate how complex is the interaction of Ascaris with the host immune system, especially, in the context of reinfections, co-infections and associated co-morbidities.

Sources of information about gestational weight gain, diet and exercise among Brazilian immigrant women living in the USA: a cross-sectional study.

OBJECTIVE: The objective of this study was to assess sources of information about gestational weight gain (GWG), diet and exercise among first-time pregnant Brazilian women in the USA. DESIGN: Cross-sectional survey. SETTING: Massachusetts, USA. PARTICIPANTS: First-time pregnant Brazilian women. RESULTS: Eighty-six women, the majority of whom were immigrants (96·5 %) classified as having low acculturation levels (68 %), participated in the study. Approximately two-thirds of respondents had sought information about GWG (72·1 %), diet (79·1 %) and exercise (74·4 %) via the internet. Women classified as having low acculturation levels were more likely to seek information about GWG via the internet (OR = 7·55; 95 % CI 1·41, 40·26) than those with high acculturation levels after adjusting for age and receiving information about GWG from healthcare provider (doctor or midwife). Moreover, many respondents reported seeking information about GWG (67 %), diet (71 %) and exercise (52 %) from family and friends. Women who self-identified as being overweight pre-pregnancy were less likely to seek information about diet (OR = 0·32; 95 % CI 0·11, 0·93) and exercise (OR = 0·33; 95 % CI 0·11, 0·96) from family and friends than those who self-identified being normal-weight pre-pregnancy. CONCLUSIONS: This is the first study to assess sources of information about GWG, diet and exercise among pregnant Brazilian immigrants in the USA. Findings have implications for the design of interventions and suggest the potential of mHealth intervention as low-cost, easy access option for delivering culturally and linguistically tailored evidence-based information about GWG incorporating behavioural change practices to this growing immigrant group.

Persistent Mullerian duct Syndrome in a Brazilian miniature schnauzer dog.

Here we describe an eight-year-old miniature schnauzer (MS) dog from Brazil with Persistent Mullerian Duct Syndrome (PMDS) and the single base pair substitution in AMHR2 exon 3, first detected in this breed in the USA. This finding is evidence of mutation dissemination to South America. In PMDS, a type of XY Disorder of Sex Development (DSD), dogs with a male karyotype and external phenotype also have a uterus, oviducts, and a cranial vagina internally. Approximately half of PMDS MS are unilaterally or bilaterally cryptorchid and many develop pyometra and/or Sertoli cell tumor. Bilateral Sertoli cell testicular tumors were present in this case, and the dog died a few days after surgical castration and hysterectomy. Although the karyotype was compatible with male chromosomal sex, a Robertsonian translocation was also identified, which may be an incidental finding. This report emphasizes the importance of cytogenetic and molecular genetic analyses, along with clinical examination, to identify chromosomal or genetic variations associated with canine PMDS. These are helpful tools to obtain early diagnosis in the MS, which is important to improve health outcomes for affected dogs and to reduce the prevalence of PMDS and cryptorchidism in this breed by avoiding the mating of carriers.

The use of microsatellite markers in Neotropical studies of wild birds: a literature review.

Despite extensive habitat fragmentation, the Neotropical region possesses 30% of the world´s bird species. Microsatellites have remained one of the most popular genetic markers and have been used in ecological and conservation studies since the 1990's. We conducted a literature review comparing the number of papers published from January 1990 to July 2015 that used microsatellite markers for studies of wild birds in the Neotropical region, USA and some European countries. We assigned the articles to three categories of studies: population genetics, animal behavior/kinship analysis and the development of species-specific bird microsatellite markers. We also compared the studies in the Neotropics that used heterologous versus species-specific markers and provide a list of heterologous markers of utility in multiple birds. Despite the rich bird fauna in the Neotropics, the number of articles published represents only 5.6% of that published by the USA and selected European countries. Within the Neotropical region, Brazil possessed 60.5% of the total papers published, with the remaining 39.5% shared between five countries. We conclude that the lack of specialized laboratories and resources still represents a limit to microsatellite-based genetic studies of birds within the Neotropical region. To overcome these limitations, we suggest the use of heterologous microsatellite markers as a cost-effective and time-effective tool to assist ecological studies of wild birds.

Cytogenetics and Molecular Genetic Analysis of Chimerism in Marmosets (Callithrix: Primates).

The birth of fraternal twins is a characteristic frequently observed in callitrichids. Cytogenetic studies have demonstrated hematopoietic chimerism in marmosets with the occurrence of two cell lines 2n=46,XX/46,XY in females and males co-twins, without phenotypic changes. Amplification by PCR have also been used to verify the presence of the SRY gene in female chimaeras. Our aim was to verify the occurrence of chimerism in Callithrix sp. individuals considered as hybrids according to their intermediate phenotypes between C. jacchus and C. penicillata. Blood samples from 37 Callithrix sp. individuals were collected. Hematopoietic chimerism 2n=46,XX/46,XY was detected by cytogenetic analysis in five individuals, three males and two females. A fragment of approximately 200bp of the SRY gene was amplified in seven females with normal external genitalia. The percentage of 32% of chimeric individuals detected in the present study is similar to that observed for pure specimens of Callithrix. These data suggests that hybridization probably does not interfere with the occurrence of twin gestation, nor of chimerism. Although cytogenetics is the main tool to identify the two cell lineages present in cases of chimerism, the amplification of the SRY gene by PCR has proved to be more efficient to identify the Y chromosome in cases of chimeric female marmoset.

Coverage and determinants of childhood vaccination during the COVID-19 pandemic in Fortaleza, Northeastern Brazil: a longitudinal analysis.

Brazil has seen a decrease in vaccination coverage since 2016. This study analyzes the immunization status of children born during the COVID-19 pandemic in Fortaleza, Northeastern Brazil. This is a longitudinal analysis that included vaccination data of 313 children aged 12 and 18 months. Vaccination cards were checked for dose application considering the schedule of immunization recommended by the Brazilian Ministry of Health. Factors associated with no retention of vaccination cards and incomplete immunization by 18 months were identified by Tobit regression analysis. About 73% of mothers presented their child's vaccination card. Non-availability of vaccination cards was associated with maternal age < 25 years and mothers with paid jobs. Only 33% and 45% of the children aged 12 and 18 months had all vaccines up to date, respectively. For 3-dose vaccines, the delay rate was around 10% for the first dose application, but 40% for the third dose. Despite delays, most children with available vaccine cards had coverage above 90% by 18 months of age. Adjusted factors associated with incomplete vaccination included living in a household with more than one child (p = 0.010) and monthly income of less than one minimum wage (p = 0.006). Therefore, delays in child vaccine application were high during the COVID-19 pandemic but a considerable uptake by 18 months of age was found. Poorer families with more than one child were particularly at risk of not fully immunizing their children and should be the target of public policies.

Factors associated with small ruminant lentivirus infection in goat herds from Pernambuco state, Northeast region of Brazil.

Serum samples (n = 1532) were collected between May 2011 to April 2012 from goats from 76 herds (49 from dairy farms and 27 herds for genetic improvement) from three geographical regions from the state of Pernambuco, Brazil: Zona da Mata, Agreste, and Sertão. Samples were processed using agar gel immunodiffusion test, with p28 CAEV antigen. The objective was to determine the risk factors for small ruminant lentivirus (SRLV) in dairy goats and goats with high genetic value. Overall, seroprevalence was 13.7% (210/1532) [95% CI: 12-15.4%] in animals and 67.1% (51/76) [95% CI: 56.5%- 77.7%] in herds. In dairy farms the seroprevalence was 73.5% (36/49) [95% CI: 61.1%- 85.8%], and in properties with animals of high genetic value it was 55.6% (15/27) [95% CI: 36.8%- 74.3%]. Robust Poisson regression analysis adjusted by the random effect of the herd showed that risk factors were: importing bucks from another Brazilian state (prevalence ratio [PR] = 4.73 [95% CI: 2.05; 10.88]), not isolating sick animals (PR = 3.27 [95% CI: 2.24; 4.76]), and participating in fairs/animal crowding (PR = 1.52 [95% CI: 1.09; 2.11]). Prevalence results show that SRLV is present in caprine herds in the state of Pernambuco and identified risk factors are strongly related to animal transit. Considering the epidemiological situation, the first step for mitigating the consequences of this disease would be controlling animal transit.

Occurrence of IgG antibodies against Toxoplasma gondii, Neospora caninum, and Leptospira spp. in goats and sheep from an indigenous village in Pernambuco, Brazil.

This study aimed to determine the occurrence of anti-Toxoplasma gondii, Neospora caninum, and Leptospira spp. antibodies in sheep and goats raised in villages of the Xukuru do Ororubá indigenous community, Pernambuco, Brazil. A total of 180 serum samples from sheep and 108 serum samples from goats of both sexes and different ages were analyzed. For antibody research, indirect immunofluorescence antibody test (IFAT) were used for the protozoa T. gondii and N. caninum, and microscopic agglutination test (MAT) for Leptospira spp., with a cutoff titer of 1:64, 1:50 and 1:100, respectively. The frequency of anti-T. gondii antibodies was 16.6% (30/180) for sheep and 11.1% (12/108) for goats. The frequency of anti-N. caninum antibodies was 10.55% (19/180) for sheep, and 20.37% (22/108) for goats, while for Leptospira spp., 2.2% (4/180) of sheep and 1.85% (2/108) of goats reacted positively. The results obtained in this study are unprecedented in indigenous communities in the country and serve as an alert for monitoring goats and sheep from the Xukuru do Ororubá indigenous village regarding the occurrence and productive impact of infections by T. gondii, N. caninum, and Leptospira spp., in addition to the occurrence of the zoonosis toxoplasmosis and leptospirosis in the indigenous community.

Bioaccessibility and oral immunization efficacy of a chimeric protein vaccine against Ascaris suum.

Human ascariasis has been characterized as the most prevalent neglected tropical disease worldwide. There is an urgent need for search to alternative prevention and control methods for ascariasis. Here we aimed to establish a protocol of oral immunization with a previously described chimera protein capable of resist through digestion and induce mucous protection against Ascaris suum infection. Mice were oral immunized with seven doses with one day interval and challenged with A. suum ten days after the last dose. In vitro digestion showed that 64% of chimeric protein was bioaccessible for absorption after digestion. Immunized mice display 66,2% reduction of larval burden in lungs compared to control group. In conclusion we demonstrated that oral immunization with chimera protein protects the host against A. suum larval migration leading to less pronounced histopathological lesions.

Beta-Carotene Supplementation for Sows: Reproductive Parameters and Productive Performance of Piglets.

The rapid fetal development and the increased demand for milk result in a catabolic state and oxidative stress in hyperprolific sows. Despite animal defense mechanisms, the dietary supplementation of antioxidants is being evaluated to reduce the impacts of excess free radicals. The aim of this study was to evaluate the short- and long-term effects of beta-carotene supplementation for sows on the reproductive response and performance of suckling piglets. A total of 120 sows were distributed in a 3 × 4 factorial arrangement of three supplementation levels (B0-no supplementation, B200-200 mg beta-carotene/day and B400-400 mg beta-carotene/day) and four groups of parity order (1st, 2nd, 3rd, above 4th). Beta-carotene supplementation during lactation resulted in a higher litter weight at weaning. A higher average weight of piglets and litter weight at birth were observed, as well as a greater number of piglets weaned and litter weight at weaning in sows supplemented with 400 mg of beta-carotene during gestation and lactation periods. Supplementation with 200 and 400 mg of beta-carotene resulted in a greater weight at weaning and daily weight gain in piglets. Daily supplementation with 400 mg of beta-carotene in the prepartum and lactation phases provides a greater litter weight at weaning and, when supplemented in the pre-gestation and gestation periods, results in a greater litter weight at birth and at weaning.

Bovine Leptospirosis in Caatinga Biome, Brazil: New Insights into Diagnosis and Epidemiology.

Bovine leptospirosis causes economic losses and raises public health concerns. It is possible that there are peculiarities in the epidemiology of leptospirosis in regions with a semiarid climate, such as the Caatinga biome in Brazil, where the climate is hot and dry, and the etiological agent require alternative routes of transmission. This study aimed to close knowledge gaps to the diagnosis and epidemiology of Leptospira spp. infection in cows from the Caatinga biome, Brazil. Samples of the blood, urinary tract (urine, bladder and kidney) and reproductive tract (vaginal fluid, uterus, uterine tube, ovary and placenta) were collected from 42 slaughtered cows. Diagnostic tests included were the microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Anti-Leptospira spp. antibodies were found in 27 (64.3%) of the animals analyzed using MAT at a 1:50 dilution (cut-off 50), while 31 (73.8%) animals had at least one organ/fluid where the presence of Leptospira spp. DNA was identified, and 29 animals (69%) were positive at bacteriological culture. The highest sensitivity values for MAT were obtained at the cut-off point of 50. In conclusion, even under hot and dry climate conditions, it is possible that Leptospira spp. can spread through alternative routes such as venereal transmission; moreover, a cut-off of 50 is recommended for the serological diagnosis of cattle from the Caatinga biome.

Healthcare Providers' Advice About Gestational Weight Gain, Diet, and Exercise: a Cross-Sectional Study with Brazilian Immigrant Women in the USA.

This study sought to quantify healthcare providers' advice about gestational weight gain (GWG), diet, and exercise among first-time pregnant Brazilian women living in the USA. This is a cross-sectional study of first-time pregnant Brazilian women living in selected cities in Massachusetts. The study sample included 86 women, nearly all (97%; n = 83) immigrants. Participants' mean age was 28.3 years and mean gestation was 27.5 weeks. Approximately 25.6% (n = 22) reported being overweight, and 1.2% (n = 1) being underweight. Only about 62% (n = 53) reported receiving advice from their healthcare provider on recommended GWG. In addition, about 83% (n = 71) and 81% (n = 70) reported receiving healthcare providers' advice on exercise and diet, respectively. Women who self-reported being overweight pre-pregnancy (OR 0.15; 95% confidence interval [CI] 0.03-0.73; p = 0.02) and women who were classified low acculturation levels (OR 0.12; 95% CI 0.02-0.08; p = 0.03) were less likely to report receiving healthcare providers' advice on GWG compared to women who self-reported being normal weight pre-pregnancy and those classified having high acculturation level, respectively. Furthermore, women who self-reported being overweight pre-pregnancy were less likely to report receiving diet-related advice (OR 0.32; 95% CI 0.10-1.02; p = 0.05) than women who self-reported being normal weight pre-pregnancy. Findings indicate the need for increased communication and counseling between healthcare providers and Brazilian immigrant women about GWG, in particular for women with low acculturation levels and language barriers suggesting the need for linguistic and culturally relevant interventions designed to improve Brazilian immigrant women's access to evidence-based information about GWG, diet, and exercise.

Global Prevalence of RNA-Positive Horses for Hepacivirus (EqHV): Systematic Review and Meta-Analysis.

Equine hepacivirus (EqHV) belongs to the Flaviviridae family, genus Hepacivirus and has the greatest genomic identity with the hepatitis C virus (HCV), one of the main causes of chronic liver disease in humans. Due to the limited applicability of studies of HCV in animal hosts, the interest in studies of characterization of viral homologues has been growing. For this reason, we performed a systematic review of the literature with meta-analysis of the prevalence of EqHV and genetic sequencing studies. Twenty-three studies from four different continents were analyzed. The OR meta-analysis (0.98; 95% CI = 0.69-1.39) showed no influence of sex (female or male) on the risk of infection. Variables associated with EqHV infection were indirectly related to animal management such as transport, reproductive practices, among others. The combined prevalence of positive animals was 7.88% (95% CI = 5.23-11.69%), with the highest proportions in Asia (16.13%; 95% CI = 7.79-30.43%), followed by South America (12.03%; 95% CI = 9.58-15.01%), Africa (8.69%; 95% CI = 6.71-11.20%), and Europe (3.63%; 95% CI = 2.10-6.22%). However, these results represent the regional stratification of the epidemiological studies for EqHV published to date and, therefore, cannot extrapolate to determine the continental prevalence of EqHV. It is therefore important to update the systematic review as further research becomes available.

Primary Health Care during the COVID-19 pandemic in Fortaleza, Brazil: associated factors and pattern of use by mothers and children up to 18 months of age.

OBJECTIVE: To analyze the delay or failure to seek primary health care by the mother-child dyads during the COVID-19 pandemic, a practice that has a high potential to increase maternal and child morbidity and mortality. METHODS: Data from three survey rounds of the Iracema-COVID cohort study, collected 6, 12, and 18 months after birth, showed the patterns of postpartum attendance to primary health care consultation of the mother-child dyad. Crude and adjusted multinomial logistic regressions with robust variance were used to assess factors associated with nonattendance. RESULTS: Among the 314 cohort mothers, 25% did not attend any primary health care consultation during the 18-months postpartum, while 30% of the mothers did all three. Regarding the child, 75% had regular primary health care consultations in all three survey rounds, while 4% did not attend any in their first 18 months of life. By the end of the first COVID-19 wave, the proportion of mother and child who attended the consultations had fallen by 23 and 18%, respectively. The main factors associated with nonattendance were mothers aged below 25 years, and mothers with more than one child. CONCLUSION: An important delay or nonattendance to primary health care consultation by the mother-child dyad was observed during the COVID-19 pandemic. Such practice, with a high potential to increase maternal and child morbidity and mortality, was particularly frequent among younger mothers and those with more than one child.

New insights on Leptospira sp. infection in ewes maintained in field semiarid conditions.

Leptospirosis is poorly studied in small ruminants raised in field semiarid conditions. In this study we compared serological, bacteriological and molecular diagnostic methods in ewes maintained in field Brazilian semiarid conditions. Blood, vaginal fluid and urine samples were collected from 60 Morada Nova ewes raised in a semi-intensive system in the Brazilian semiarid. Diagnostic tests performed were microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Anti-Leptospira sp. antibodies were found in eight (13.33%) animals analyzed by MAT at reciprocal titer 25 (cut-off 25), while Leptospira sp. DNA was detected in urine or vaginal fluid of 56 animals (93.33%). There was growth of leptospires in 10 urine cultures and in 11 vaginal fluid cultures, however, two of urine (2/60-3.33%) and eight cultures of vaginal fluid (8/60-13.33%) were confirmed by PCR. Two samples of vaginal fluid (one of each animal) were submitted to sequencing demonstrating 99% similarity with L. santarosai and L. interrogans. The highest MAT sensitivities were obtained with reciprocal titer 25 (cut-off 25) compared to 50 and 100. The performance of different diagnostic techniques for leptospirosis in ewes raised in field semiarid conditions allowed a better evaluation of the herd, as well as made it possible to identify carrier animals. Genital route may be important for efficient transmission and without dependence on environmental factors in ewes from semiarid, as well as it's highlighted that titer 1:25 in serology was more efficient, indicating its use in ewes in field semiarid conditions.

Uniparental genetic markers to investigate hybridization in wild-born marmosets with a mixed phenotype among Callithrix aurita and invasive species.

The native marmoset of the Southeastern Atlantic Forest in Brazil is among the 25 most endangered primates of the world. Hybridization with alien species is one of its main threats registered since the early 2000s based on phenotype, so far, without genetic confirmation. Using uniparental molecular markers, we analyzed 18 putative hybrids, captured from 2004 to 2013 in different localities of the Atlantic Forest. A nine base pair deletion in the SRY gene of C. aurita was used to investigate paternal ancestry. Maternal ancestry was assessed by DNA sequencing of ca. 455 bp from the COX2 gene. Hybridization was confirmed for 16 out of the 18 marmosets since they inherited COX2 haplotypes of the alien C. penicillata or C. jacchus and the SRY deletion specific to C. aurita. Two individuals inherited both parental lineages of C. aurita, which is probably related to backcrossing or hybrid interbreeding. The direction of hybridization of females with the matrilineal lineage of invasive species with males descending from the native lineage was predominant in our sampling. This is the first time that hybridization between C. aurita and invasive species has been confirmed through genetic analysis.