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Brazil introduced rotavirus vaccination in March 2006. We studied 133 rotavirus-positive fecal samples collected from February 2005 through December 2007. Genotype G2P[4] was found in 1.4% of samples in 2005, in 44% in 2006, and in 96% in 2007. Rotavirus detection rate decreased from 38% in 2005 to 24% in 2007 (p = 0.012). [ABSTRACT FROM AUTHOR]

Bovine genital leptospirosis (BGL) is a chronic reproductive disease in cattle, often causing significant economic losses, and is commonly associated with leptospiral strains belonging to the Sejroe serogroup. A two-step protocol was recommended, based on serological screening of the herds followed by an individual diagnosis with PCR of a genital sample. Although proposed, it has not been commonly applied under field conditions, leading to frustrating outcomes in disease control. In that context, the present study aimed to demonstrate the viability of that two-step protocol under field conditions for diagnosing BGL in eight herds with reproductive disorders. Blood samples were collected from 440 cows for serology. In addition, 304 cervicovaginal mucus (CVM) samples were collected for lip L32-PCR and 11 samples were sequenced of the sec Y gene. All herds showed high seroreactivity, mainly against the serogroup Sejroe. In addition, 113 of the 304 CVM samples (37.2%) were PCR-positive. DNA sequencing of 11 positive samples based on the sec Y gene revealed maximum identity (100%) with L. interrogans species. This study found CVM sampling quick and easy, making it practical for field use. Overall, the results support the two-step protocol serological screening followed by CVM-PCR testing as an efficient and reliable method for diagnosing BGL in herds with reproductive disorders in field conditions.

Pathogenic bacteria Leptospira spp. are commonly associated with bovine leptospirosis, characterized chiefly by chronic and subclinical reproductive disorders. Strains from the Sejroe serogroup play a significant role in these chronic genital infections known as Bovine Genital Leptospirosis (BGL), which notably impact cattle health. This study aims to deepen our understanding of BGL by investigating the genetic diversity, geographical distribution, and specific anatomical sites of infection of the causative agents. Initially, uterine fragments and cervicovaginal mucus were collected from 47 cows and subjected to PCR targeting the lipL32 gene. Positive samples in lipL32-PCR (9 samples) underwent genotyping based on the secY gene. Subsequently, sequences were aligned with GenBank entries (108 sequences) and analyzed in silico. All nine sequences from this study were identified as L. interrogans with an identity >99 % to serogroup Sejroe reference strains (Norma and L53). In the broader analysis, the most prevalent species observed was L. borgpetersenii, followed by L. interrogans and L. santarosai. The haplotype network of L. interrogans revealed that haplogroups B and C exclusively included L. interrogans strains of genital origin, while haplogroup A encompassed strains from renal sources as well. These findings underscore the significance of the L. borgpetersenii genotype Hardjobovis and L. interrogans genotype Hardjoprajitno as the predominant circulating strains and highlight the existence of distinct haplogroups of pathogenic leptospires originating from genital sources. We advocate for the use of secY as an effective genetic marker for Leptospira spp. and stress the necessity for additional research prioritizing the genital tract. The outcomes of this study contribute to the development of improved control measures for chronic cattle diseases and provide valuable guidance for future investigations., Competing Interests: Declaration of Competing Interest None of the authors has any other financial or personal relationships that could inappropriately influence or bias the content of the paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Bovine leptospirosis is a worldwide disease that causes reproductive issues, including early embryonic death, stillbirths and infertility, which result in significant economic losses. Although bovine leptospirosis is well-documented in cows, the role of bulls in harbouring and potentially transmitting genital leptospirosis has been largely neglected. We examined genital and blood samples from 16 slaughtered bulls using microscopic agglutination testing (MAT), polymerase chain reaction (PCR) and sequencing of amplicons. Our results showed that 81.2% of bulls were seroreactive, and 75% were genitally infected. The amplicons displayed an identity greater than 99% with Leptospira interrogans strains from the Sejroe serogroup, specifically serovar Hardjoprajitno. This study demonstrates that bulls can harbour in their genital tract the same leptospires associated with reproductive syndromes in cows from the same geographic region, highlighting the importance of bulls in maintaining and transmitting Sejroe serogroup strains associated with reproductive disease., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Background: Viral hepatitis is a disease that is more prevalent among individuals residing in remote regions and in contexts of social vulnerability. The objective of this study was to ascertain the seroprevalence of hepatitis A (HAV), B (HBV), and C (HCV) in vulnerable communities in the rural area of São João do Piauí (SJP), northern Brazil., Methods: Immunoenzymatic assays were employed to detect the presence of anti-HAV (total and IgM), HBsAg, anti-HBc, anti-HBs, and anti-HCV serological markers in serum samples. Samples that yielded positive results were subjected to further analysis using real-time polymerase chain reaction (PCR)., Results: A total of 492 individuals, ranging in age from 3 to 101 years, were consecutively recruited from eight rural communities. The majority of individuals were female (51.2%), over 30 years of age (57.1%), self-identified as Black/Brown (92.2%), and resided in Afro-Brazilian communities, designated as "quilombos" (68.1%). The seroprevalence of anti-HAV was 69.5% (95% CI: 65.4-73.6%), while that of anti-HBc was 4.7% (95% CI: 2.8-6.6%), and that of anti-HBs was 35.2% (95% CI: 30.1-39.4%), and 0.2% (95% CI:0.0-0.6%) for anti-HCV., Conclusions: The seroprevalence rates observed were higher than the national average, and a significant proportion of individuals in the target age groups were susceptible to HBV, despite the availability of vaccination. These findings highlight potential shortcomings in the management of vaccine-preventable diseases, which could have implications for public health.

The reproductive features of equine leptospirosis are often neglected. Equine genital leptospirosis is characterized as a silent chronic syndrome, and besides abortions, leads to placental abnormalities, stillbirths, and birth of weak foals. This study aimed to study the occurrence of placental abnormalities associated with Leptospira interrogans infection in naturally infected mares under field conditions. The studied herd had a high occurrence of placentitis and abortions. Ten pregnant mares, eight with placental abnormalities on ultrasonography and were selected. Serum and cervicovaginal mucus (CVM) samples were collected for serology and PCR, respectively. Positive samples in lipL32-PCR were submitted to the sequencing of the secY gene. In lipL32-PCR of CVM, five out of 10 (50%) mares were positive and all were characterized as Leptospira interrogans. Our results highlight the presence of placental abnormalities in the reproductive subclinical leptospirosis syndrome. We encourage field veterinarians to include leptospirosis testing in their reproductive management., Competing Interests: Declaration of competing interest None of the authors has any financial or personal relationships that could inappropriately influence or bias the content of the paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Sandflies are known vectors of leishmaniasis. In the Old World, sandflies are also vectors of viruses while little is known about the capacity of New World insects to transmit viruses to humans. Here, we relate the identification of RNA sequences with homology to rhabdovirus nucleocapsids (NcPs) genes, initially in the Lutzomyia longipalpis LL5 cell lineage, named NcP1.1 and NcP2. The Rhabdoviridae family never retrotranscribes its RNA genome to DNA. The sequences here described were identified in cDNA and DNA from LL-5 cells and in adult insects indicating that they are transcribed endogenous viral elements (EVEs). The presence of NcP1.1 and NcP2 in the L. longipalpis genome was confirmed in silico. In addition to showing the genomic location of NcP1.1 and NcP2, we identified another rhabdoviral insertion named NcP1.2. Analysis of small RNA molecules derived from these sequences showed that NcP1.1 and NcP1.2 present a profile consistent with elements targeted by primary piRNAs, while NcP2 was restricted to the degradation profile. The presence of NcP1.1 and NcP2 was investigated in sandfly populations from South America and the Old World. These EVEs are shared by different sandfly populations in South America while none of the Old World species studied presented the insertions., Competing Interests: The authors declare no conflicts of interest.

The single-dose protocol of streptomycin treatment has been recommended to treat renal leptospirosis in bovines. However, treating genital infection remains a challenge. Recently, a protocol using three doses of streptomycin demonstrated effectiveness in the genital clearance of experimentally infected ewes. Therefore, the present study aimed to apply this three-dose protocol for genital infection treatment in naturally infected cows under field conditions. Thirty beef cows were diagnosed as positive by lipL32-PCR in their genital samples. Nucleotide sequences (n = 10) characterized them as Leptospira interrogans sg Sejroe, genetically related to Hardjoprajitno strains. After molecular diagnosis, 13 cows received a single dose of 25 mg/kg streptomycin. The other 17 cows were submitted to the three-dose protocol. The successful treatment rate of genital infection on the single streptomycin dose was 7/13 (53.8%), while the cows that received the three doses 16/17 were negative (94.1% of efficacy). Based on those results, we conclude that the standard treatment preconized for renal infection is not adequate for genital infection, and the three-dose protocol was successful in eliminating the carrier status of genital leptospirosis., Competing Interests: Declaration of Competing Interest We have no conflict of interest to declare., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The aim of the present study was to assess morphologic and genetic data on ascariasis in swine (Sus scrofa domesticus) and humans in low-resource rural and periurban communities in the state of Piauí, Brazil. Our cross-sectional survey included 100 fecal samples obtained from swine and 682 samples from humans. Fifteen pigs were necropsied. Human and porcine fecal samples were examined to identify Ascaris eggs. Parasites obtained in the swine necropsies were studied using scanning electron microscopy (SEM), and the mitochondrial gene encoding the cytochrome oxidase 1 (cox1) enzyme was partially amplified and sequenced for molecular taxonomy and phylogenetic analyses. The overall prevalence of Ascaris eggs in the swine fecal samples was 16/100 (16%). No Ascaris eggs were identified in the human fecal samples. SEM of six worms recovered from pigs demonstrated morphological characteristics of A. suum. Cox1 sequences were compatible with A. suum reference sequences. Original and reference (GenBank) nucleotide sequences were organized into clusters that did not segregate the parasites by host species or and region. The largest haplogroups were dominated by haplotypes H01, H02 and H31. In the communities studied, there was no epidemiological evidence of the zoonotic transmission of ascariasis at the human-swine interface.

This study aimed to assess the factors associated with mucosal leishmaniasis (ML) within the scope of tegumentary leishmaniasis (TL) cases reported in Brazil. Surveillance data were assessed, and comparisons were made between ML and cutaneous leishmaniasis (CL) cases. Additionally, ML incidence rates for municipalities were depicted through a geographic information system. From 2007 to 2017, 235,489 TL cases were reported, of which 235,232 were classified as follows: 14,204 (6%) were ML cases and 221,028 (94%) were CL cases. Multivariate analysis showed that the proportion of ML cases reached 16.8% among individuals >75 years (adjusted OR = 2.77; 95% CI = 2.41-3.19; p < 0.001), and ML was also more frequent among males (aOR = 1.28; 95% CI = 1.20-1.38; p < 0.001), HIV-positive patients (aOR = 2.15; 95% CI = 1.80-2.56; p < 0.001), patients residing in urban areas (aOR = 1.52; 95% CI = 1.43-1.62; p < 0.001), and imported cases (with respect to county) when compared to autochthonous cases (aOR = 1.84; 95% CI = 1.71-1.98; p < 0.001). A lower proportion of positive results in direct parasitological examinations was observed in ML cases (32.6% vs. 60.8%; p < 0.001). The leishmanin skin test results were more often positive in ML cases (41.7% vs. 25.9%; p < 0.001). In ML, compatible changes in histopathology were more frequent (14.6% vs. 3.9%; p < 0.001). A greater proportion of ML cases were treated with amphotericin B (6.9% vs. 0.9%; p < 0.001). The case-fatality rate was higher in ML (0.6% vs. 0.1%; p < 0.001). A higher incidence of ML was observed in a geographical band extending across the Amazon region from the southern Para State to the Acre State. ML exhibited varying frequencies within specific populations. The definition of predictable factors predisposing Leishmania-infected subjects to develop ML is important for defining strategies to mitigate the mucosal damage caused by leishmaniasis.

Background: Bovine leptospirosis is an important reproductive disease and abortion is a major sign, leading to economic impacts. Due to its multifactorial etiology, the proper diagnosis of the cause of the abortion is crucial. Necropsy of the fetuses followed by molecular analysis is recommended for diagnosis, and the investigation mainly occurs in the kidneys and liver. This study aimed to analyze unconventional sites for the presence of leptospiral DNA in bovine anicteric aborted fetuses., Methods: Five fetuses of the same herd were received for necropsy and diagnosis. Conventional lip L32-PCR was performed in the fetuses' kidneys, livers, lungs, hearts, spleens, subcapsular kidney content, abomasal fluid, and in the cavity's hemorrhagic contents. To complete the investigation, the sera of 30 cows of the herd were collected to perform the serologic screening by Microscopic Agglutination Test. In addition, six subfertile non-pregnant cows from the same herd were selected due to their low reproductive performance, and genital samples (uterine fragment and cervicovaginal mucus) and urine were collected for lip L32-PCR. PCR-positive samples were submitted to a nested PCR of the sec Y gene and intended for sequencing., Results: The herd presented seroreactive animals (11/30, 36.6%), all against the Sejroe serogroup, with titers between 200 and 1600. In necropsy, four fetuses showed hemorrhagic and anicteric lesions, while one fetus had no macroscopic lesions. Regarding molecular analysis, all the fetuses were positive in lip L32-PCR and the positive sites were the heart, lungs, subcapsular kidney content, thymus, kidneys, liver, and abomasal fluid. Only one fetus presented positive results in the kidney and liver, while three fetuses were positive in the abomasal fluid. Five of six cows were positive for lip L32-PCR, all being positive only in genital samples. Of the fetuses and the cows, seven sequences were obtained and all were identified as Leptospira interrogans serogroup Sejroe serovar Hardjoprajitno., Conclusions: In order to improve the diagnosis of leptospirosis in cows, it is recommended to perform a comprehensive analysis of the samples, beyond the kidneys and liver. Thus, we highly encourage testing multiple organs by PCR to investigate abortions suspected of bovine leptospirosis, particularly in anicteric fetuses.

Background: Intestinal parasite Giardia can affect children's physical development mainly stunting even in asymptomatic cases. The protozoa G. lamblia is divided into assemblages A-H. However, it is still unclear whether clinical manifestations and pathogenesis may vary according to the infecting assemblage., Objectives: To investigate whether G. lamblia assemblages influence differently the physical development of preschoolers from a community of Rio de Janeiro, Brazil., Methods: Anthropometric parameters were analysed from children attending a daycare centre and stool samples were obtained for the G. lamblia diagnosis. G. lamblia isolates from positive samples were genotyped. Data were analysed in order to verify whether there is a relationship between G. lamblia infection and the physical development of children according to the assemblage., Findings: Herein we demonstrated that although eutrophic, G. lamblia-infected daycare preschoolers from a low-income community presented growth delay compared to non-infected ones. This effect was observed for the three assemblages (A, B or E) found infecting humans., Main Conclusion: G. lamblia causes growth delays on children independent of infecting assemblage (A, B or E).

Bovine leptospirosis has as main causative agents Leptospira spp. from Sejroe serogroup. Vaccination is a crucial step to control this infection. The use of conserved proteins among Leptospira spp. is of great importance for a protective immune response. The aim of the present study is to genetically analyze antigens of Leptospira spp. from Sejroe serogroup strains isolated from cattle for a preliminary evaluation of vaccine candidates. Genes associated with antigenicity-LigA, LipL32, Loa22, and OmpL1-were analyzed through bioinformatic and immunoinformatic tools. Despite high diversity observed in strains, on an amino acid level, highly conserved regions were observed (> 90%), particularly in LipL32 gene. Moreover, highly conserved amino acid regions (> 30 aa) were observed in all genes, regardless of species, geographical origin or biological source of isolation. Superposed structures of protein fragments including all the predicted MHC-II and B-Cell epitopes were demonstrated. Results presented herein are preliminary, but a fundamental step towards the development of an efficient vaccine against bovine leptospirosis, a silent but enormously concerning disease., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: The expansion of rotavirus (RV) immunization in several countries reduced the burden of acute diarrheal disease (ADD) and diarrhea-associated mortality. Although community transmission of live attenuated monovalent rotavirus vaccine (G1P[8] RV1) virus has been demonstrated in children and household contacts, fecal shedding of these strains in neonates and infants under six weeks of age has never been demonstrated. The objective of the study was to assess ADD and rotavirus vaccine strain shedding before and after immunization through 24 months of age., Methods: This was a prospective cohort study in a low-resource community in which stool samples were collected from neonates from 15 to 45 days of age every 2 weeks, after both doses of G1P[8] RV1, and in subsequent ADD episodes until 2 years of age. RV was detected and genotyped in stool samples by RT-PCR., Results: We enrolled 242 participants who were followed for an average of 23 months. The specific prevalence of G1P[8] RV1 virus was 3.3% in neonates and infants less than six weeks of age, 50% after the first dose, and 25.6% after the second dose. Among the 70 participants with ADD, G1P[8] RV1 virus was identified in only one participant (1.4% prevalence)., Conclusions: In vaccinated children, there were no breakthrough infections with G1P[8] RV1 and ADD was rare supporting high vaccine effectiveness. We observed G1P[8] RV1 virus shedding among neonates and infants before the first vaccine dose, providing evidence of transmission of the vaccine strain from immunized children to those who are not yet vaccinated., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)

Leptospirosis is a bacterial disease caused by the infection of pathogenic strains of the genus Leptospira, endemic in tropical and subtropical regions. Although well documented in terrestrial animals and humans, little information is available on its distribution and impact on marine animals. Despite clinical manifestations that may occur, the occurrence of carriers was suggested in some species. Nevertheless, there are few studies regarding the infection by Leptospira sp. in marine mammals. In this context, and considering the One Health approach, the present aimed to investigate pinnipeds' role as Leptospira sp. carriers. Kidneys of 47 pinnipeds of two species, Arctocephalus australis (n = 40) and Arctocephalus tropicalis (n = 7) were collected. DNA was extracted and the diagnosis was performed through LipL32-PCR and genetic characterization based on secY gene sequencing. Phylogenetic analysis and haplotype networks were constructed. Pathogenic Leptospira sp. DNA was detected in 31.9% (15/47) of the tested pinnipeds. It was possible to amplify and sequence eight strains (6 for A. australis, 2 for A. tropicalis), all identified as L. interrogans, with high similarity with sequences from Icterohaemorrhagiae serogroup. Phylogenetic analysis revealed sequences from the present study grouped in species-specific unique clusters, but very close to others from humans, wild animals, and domestic animals. We demonstrate that pinnipeds could act as carriers, and play an important role in leptospirosis dynamics., (Copyright © 2022. Published by Elsevier Ltd.)

Bovine Genital Leptospirosis (BGL) is an important syndrome that leads to reproductive failures. The present study aimed to perform a molecular analysis of Leptospira spp. identified from genital and urine samples from in vivo naturally infected cows with poor reproductive performance. A total of 48 cows destined for culling due to low reproductive efficiency were selected and submitted to sampling. Uterine fragments, cervicovaginal mucus (CVM), and urine were collected from all of the cows and processed for culturing and PCR. One isolate was recovered from the uterus of one cow. Other 25 animals were PCR-positive, totaling 26 positive cows. Of them, 18 animals were positive in lipL32-PCR to genital samples, while only seven animals were positive in urine. From those, sequencing of secY gene was performed. Of the 21 good sequences obtained, 16 were L. interrogans, two were L. noguchii, two were L. santarosai and one was L. borgpetersenii. In order to evaluate the genetic similarity of sequences found herein and other sequences from bovines worldwide, a phylogenetic analysis and haplotype networks were performed. Cows with reproductive failures had a significant association (p < 0.05) with positive PCR of genital samples when compared to PCR of urine. None of the animals were positive for genital samples and urine simultaneously. A high diversity of leptospiral strains were found, even in animals of the same epidemiological region. Haplotype networks of L. interrogans showed clusters of sequences from the uterus and CVM with high similarity to other genital sequences originating from previous studies. L. borgpetersenii haplotype networks presented two major clusters with high similarity, even from worldwide sequences, while L. santarosai showed clusters with high genetic distances, even with all the sequences being from Brazil. This study reinforces the theory that BGL and renal infection are distinct diseases, as well as, genital samples are crucial for the diagnosis of cows with reproductive failures caused by leptospires. In addition, haplotype networks confirmed a high genetic similarity between sequences from the present study and Sejroe strains, reinforcing Sejroe strains as the main BGL agents., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Ltd.)

Background: Agile, accessible and cheap diagnosis of hepatitis C virus (HCV) infection is essential to achieve the elimination of this infection, worldwide, as mandated by the World Health Organzation as part of its strategy for 2030. Dried blood spots (DBS) can be an attractive alternative for sample collection among people living in remote areas and vulnerable populations due to the less invasive collection, its biosafety, and storage & transportation of samples at room temperature., Design: This study aims to estimate the usefulness of dried blood spot samples for the diagnosis and the assessment of HCV infection rates in three different settings in Brazil. Cross-sectional analysis of a sample collection from different populations, aiming to assess the performance of the testing algorithms and respective procedures among different populations with diverse background infection rates., Methods: We reported the evaluation of DBS as alternative samples for detecting anti-HCV in different groups in real life conditions: (I) Vulnerable subjects living in remote areas of Southeast, North and Northeast Brazil (n = 1464); (II) Beauticians (n = 288); (III) People who use non-injectable drugs (n = 201); (IV) patients referred to outpatient care (n = 275)., Results: General assay accuracy was 99%, with a weighted kappa value of 0.9, showing an excellent performance. Sensitivities ranged from 87.5% to 100.0% between groups and specificities were above 99.2%. A total of 194 individuals had HCV RNA in serum and concordance of anti-HCV detection in DBS was 98.4%., Conclusions: DBS samples could be used for anti-HCV detection in different populations recruited in real life conditions and ambulatory settings, with a high overall sensitivity and specificity., (© 2022. The Author(s).)

Leptospirosis is known to determine reproductive disorders on livestock, and Leptospira interrogans and Leptospira borgpetersenii are the most frequently reported species. Leptospira noguchii is an emerging pathogen, but its association with reproductive disease is unclear. We have detected L. noguchii as the agent of an outbreak with reproductive disorders in a Brazilian dairy goat flock. In the kidding season, five out of 10 Saanen had abortions in the final month of pregnancy and two newborn kids had acute clinical signs. After necropsy of three foetuses and one newborn kid, fragments of liver, lung and kidney were submitted to lipL32-PCR. It yielded positive results in at least one fragment from each animal. After, a nested secY-PCR, followed by sequencing, could identify L. noguchii, with 99-100% of identity with sequences obtained from cattle in the same region. For the first time, L. noguchii was detected in goats and, most importantly, the association of this leptospiral species with reproductive failures in ruminants has been demonstrated., (© 2021 Wiley-VCH GmbH.)

Some helminth species belonging to the order Strongylida are parasites of the digestive tract of vertebrates, including man and domestic animals. In humans, infections with Oesophagostomum spp. and Trichostrongylus spp. may be misdiagnosed as hookworm disease on parasitological stool examination, mainly in regions where these infections are coendemic, since eggs released in hosts' feces are morphologically similar. This study presents the morphologic characterization of Strongylida larvae recovered from humans and pigs living in close proximity, exploring putative zoonotic cycles. One hundred three humans and 27 pigs were included in low-resource rural communities in the state of Piauí, northeastern Brazil. Strongylida eggs were present in 12 (11.7%) humans and 23 (85.2%) pigs through conventional parasitological examination. Strongylida-positive fecal samples were submitted to coprocultures using the Harada-Mori technique. All 22 larvae obtained from human feces were classified as hookworms (Ancylostomatidae). From a total of 37 larvae obtained from swine, 23 (62.3%) were classified as Oesophagostomum , 6 (16.2%) were Hyostrongylus , and 4 (10.8%) were Trichostrongylus . Four larvae (10.8%) obtained from pigs were classified as Strongyloides . The morphological study of filariform larvae obtained in coprocultures is a useful and inexpensive tool in the screening of intestinal helminthiasis in a One Health approach., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2022 Polyanna A. A. Bacelar et al.)

Leptospirosis in ruminants causes reproductive failures leading to important economic losses. This study assessed the occurrence and genetically identified Leptospira spp. in the follicular fluid (FF) of naturally infected live cows. A total of 251 asymptomatic cows from different commercial dairy herds were subjected to ovum-pick up technique for follicular fluid sampling. PCR was performed for Leptospira spp. detection and phylogenetic analysis was later implemented for sequencing. From 251 samples analyzed, 67 (26.7 %) were lipL32-PCR positive, confirming the presence of leptospiral DNA on FF. Furthermore, it was possible to amplify and sequence nine strains after secY nested-PCR. All of them were identified as L. interrogans, with 100 % of identity with strains belonging to Sejroe serogroup. Our findings reveal a high occurrence of infection of Leptospira in the ovarium of asymptomatic cows, highlighting the importance of considering the silent leptospirosis syndrome when screening animals for assisted reproductive biotechniques., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Metastrongylosis is an infection of the respiratory tract of pigs caused by parasites of the genus Metastrongylus , whose eggs are similar to other Strongylida through light microscopy; species-specific identification can be performed with molecular tools. We explored the species composition and the genetic diversity of Metastrongylus infecting pigs in close contact with humans in impoverished rural communities in the state of Piauí, in northeastern Brazil. Fecal samples ( n  = 78) were collected for parasitologic tests. Egg morphometry and molecular characterization, using the cytochrome c oxidase subunit 1 ( cox 1) gene, were performed. For strongyliform eggs, 62 of 78 (80%) pigs were positive and 6 of 99 (6%) eggs had dimensions compatible with Metastrongylus . Of the 37 samples submitted to PCR, 10 were identified as M. salmi . We found 3 M. salmi haplotypes, including 2 new and 1 described previously in Europe. Overall, M. salmi demonstrated lower intraspecific genetic diversity: diversity index (H) ± SD = 0.318 ± 0.164, n  = 12, compared with published M. pudendotectus sequences (1.000 ± 0.272, n  = 3). To our knowledge, M. salmi DNA sequences have not been published previously from pigs in South America.

Bovine Genital Leptospirosis (BGL) is an important reproductive disease. The main agents are Sejroe strains, particularly the Hardjo genotypes from Leptospira interrogans and L. borgpetersenii. Although other Sejroe strain, L. santarosai genotype Guaricura, has been frequently isolated from asymptomatic and slaughtered cattle, even from vaginal fluid samples, the role of this strain as real agent of BGL remains uncertain. This study aimed to reinforce L. santarosai strain Guaricura as an important BGL agent, through genetic characterization of a uterine isolate from a live subfertile cow. Urine, cervicovaginal mucus (CVM) and uterine fragment (UF) were collected. In a set up field laboratory, urine, CVM and UF were immediately seeded in T80/40LH medium with antimicrobial cocktail STAFF. Cultures were subcultured in T80/40LH without cocktails, stored at 29ºC and weekly examined. DNA from urine, CVM and UF samples were submitted to PCR targeting lipL32 and secY genes. One leptospiral isolate was recovered from uterine sample; it was serogrouped as Sejroe (titre 25,600) and secY sequencing revealed high genetic similarity with L. santarosai strains from Guaricura serovar. The isolation of this strain from uterus of a live subfertile cow represents substantial evidence that L. santarosai strain Guaricura indeed plays an important role as a BGL agent., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Human infections with gut protozoan parasites are neglected and not targeted by specific control initiatives, leading to a knowledge gap concerning their regional diversity and epidemiology. The present study aims to explore Giardia duodenalis genetic diversity and assess the epidemiologic scenario of subclinical infections in different Brazilian biogeographic regions. Cross-sectional surveys (n = 1334 subjects) were conducted in four municipalities in order to obtain fecal samples and socioenvironmental data. Microscopy of non-diarrheal feces and nucleotide sequencing of a β-giardin gene fragment were performed. From a total of 51 samples that could be sequenced, 27 (52.9%) β-giardin sequences were characterized as assemblage A and 24 (47.1%) as assemblage B. In the Amazon, assemblage B was the most frequently detected, predominantly BIII, and with two novel sub-assemblages. Assemblage A predominated in the extra-Amazon region, with five novel sub-assemblages. Prevalence reached 17.8% (64/360) in the Amazon, 8.8% (48/544) in the Atlantic Forest, 7.4% (22/299) in Cerrado and 2.3% (3/131) in the Semiarid. People living in poverty and extreme poverty presented significantly higher positivity rates. In conclusion, subclinical giardiasis is endemic in Brazilian communities in different biogeographic regions, presenting high genetic diversity and a heterogeneous genotypic distribution.

Helminths of the genus Oesophagostomum cause enteric diseases and affect domestic animals such as pigs. The aim of this study was to explore the species composition and genetic diversity of Oesophagostomum spp. infecting pigs in close contact with humans in the state of Piauí, Brazil. Eighty-seven fecal samples were collected for parasitological tests and molecular analysis. Through microscopy, the overall positivity rate for strongyliform eggs was 81.6% among the pigs studied. Forty-two strongyliform egg samples were subjected to PCR and six cox1 sequences (637 bp) were identified for the genus Oesophagostomum. The sequences were identified as Oesophagostomum dentatum, O. quadrispinulatum and O. columbianum. In the phylogenetic tree and haplotype network, 89 sequences were separated into seven clusters, which also included reference sequences from GenBank. Oesophagostomum dentatum and O. quadrispinulatum were seen to be closely related species and formed a monophyletic group related to O. aculeatum. Oesophagostomum columbianum showed similarity with sequences from parasites infecting small ruminants and the clade was positioned closer to O. bifurcum. High interspecific diversity was found and intraspecific diversity varied according to the species. This was the first study to characterize Oesophagostomum DNA sequences obtained from pigs in Brazil.

Canine leptospirosis is a worldwide zoonosis, varying from asymptomatic and chronic infections to clinical acute disease. In many parts of the world Leptospira interrogans serogroup Icterohaemorrhagiae strains have great epidemiological importance, being the most prevalent on dogs. The present study aims to characterize and compare strains/sequences belonging to the serogroup Icterohaemorrhagiae recovered from clinically ill and asymptomatic dogs. Based on secY gene sequences of L. interrogans serovar Icterohaemorrhagiae, we have studied genetic diversity of strains obtained from 13 dogs, including dogs with clinical signs of acute leptospirosis, asymptomatic dogs and animals with chronic kidney disease, all of them from the same geographical area, the state of Rio de Janeiro, Brazil. No genetic variations on secY gene were observed between strains/groups. No significant associations were observed between clinical status and age, sex or vaccinal status. The same strain leads to different clinical outcomes on canine leptospirosis. The answer for this will rise from deep studies regarding whole genomic sequencing of the strains, as well as proteomics. Those studies may provide key information for understanding of the clinical manifestation of the disease., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The present study aimed to identify larval trematodes shed by snails found in water bodies used by urban communities in a former schistosomiasis endemic area in the state of Piauí, in the Brazilian semiarid region. A malacological survey was performed followed by analysis of the cercariae shed by the snails after light exposure. Biomphalaria straminea specimens (n=1,224) were obtained from all seven collection sites. Cercariae shed by snails were i) single tailed, in which one type of cercariae was identified ( Echinostoma cercariae), and ii) with bifurcated tail (brevifurcate apharyngeate distome, brevifurcate pharyngeate distome, and longifurcate pharyngeate distome [strigeocercaria]). Brevifurcate apharyngeate distome were further examined and the presence of spikes in swimming membranes enabled the identification of Spirorchiidae cercariae in all individuals, demonstrating the absence of cercariae compatible with Schistosoma mansoni . Nevertheless, the accurate diagnosis of S. mansoni circulation in former endemic areas is still necessary.

Swine genital leptospirosis is an infectious disease that leads to economic losses due to abortions, stillbirths, and reproductive failures. Considering the scarcity of studies regarding this condition, the objective of the present study was to identify and analyse leptospires infecting the reproductive tract of female pigs slaughtered in the Amazon region. Cervical-vaginal mucus (CVM) from 150 non-pregnant females were collected and submitted to molecular analysis. Initially, polymerase chain reaction (PCR) based on the lipL32 gene was performed. A total of 26.7% (40/150) samples were positive, indicating the presence of Leptospira sp. DNA. Subsequently, positive lipL32-PCR samples were evaluated using secY nested-PCR and sequencing procedures. Eleven amplicons could be sequenced and were identified as Leptospira interrogans (100% identity). Results from phylogenetic analyses led to identification of a putative strain of L. interrogans serogroup Australis, which is indicative of this being a serogroup. In the present study, there was detection of female pigs with leptospires in CVM indicating the possibility of venereal transmission. The large number of genital positive cases could indicate that genital leptospirosis syndrome could also be relevant onto swine production., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Chagas disease is endemic throughout most of Bolivia, with prevalence rates of 25% observed in some geographic areas located mainly in the sub-Andean region., Methods: Community-based entomological surveillance was carried out in the sub-Andean departments of Cochabamba (municipalities of Cochabamba, Punata and Sacaba), Tarija (municipality of Tarija) and Chuquisaca (municipality of Sucre). The surveillance parameters evaluated were: (i) the proportion of cards with the presence of triatomines; (ii) the distribution of positive cards by area; and (iii) the proportion of cards with the presence of infected triatomines., Results: Of the cards returned, in 852 (3.1%) there was a mention of the presence of triatomines. The species Triatoma infestans, Triatoma sordida and Triatoma guasayana were identified in 812 (95.3%), 39 (4.6%) and 1 (0.1%), respectively. The median monthly positivity rate of the cards during 2011-2018 was higher in Punata (9.1%; IQR=3.2-15.4%). The median monthly rate was highest in 2012 (2.7%; IQR=0-5.6%). Fifty positive cards (5.8%) presented insects that were positive for trypanosomatids, mainly in Cochabamba and Punata., Conclusions: The report of triatomines foci by inhabitants represents an effective surveillance system coordinated by a network of specialized and multidisciplinary health centers. These strategies, which should be included in the health policies of endemic countries, enable extending and deepening the dialogue among technicians, communities and their local authorities., (© The Author(s) 2021. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.)

Bovine genital leptospirosis (BGL) is characterized by silent chronic reproductive disorders, most related to early embryonic death leading to estrus repetition, subfertility and abortions. However, most studies were conducted in slaughterhouses, which lacks reproductive and sanitary history of the studied animals. This study aimed to evaluate the occurrence of Leptospira sp. infection in live cows with history of low reproductive efficiency. Blood, urine, cervico-vaginal mucus and uterine fragment were collected from nine cows of the same herd presenting reproductive failure (abortions, estrus repetition and chronic infertility). Serology (MAT) and molecular analysis (PCR and nucleotide sequencing) were performed. Serology showed three (33.3%) seroreactive cows, two to Sejroe and one to Icterohaemorrhagiae serogroups. Six cows (66.7%) presented leptospiral DNA on genital samples, while all urine samples were negative. L. interrogans was identified in five samples, very closely related to strains from Sejroe (n = 3) and Icterohaemorrhagiae (n = 2) serogroups, while L. noguchii was identified in one sample. Results from this preliminary study demonstrates the presence of leptospires on uterus and reinforces the negative impact of leptospiral infection on reproductive tract, highlighting its association with reproductive failures on live animals., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: This study assessed the interactions between income, nutritional status and intestinal parasitism in children in Brazil., Methods: A cross-sectional study (n = 421 children aged 1 to 14 years living in the states of Piauí (rural communities in the city of Teresina) and Rio de Janeiro (rural and periurban communities in the city of Cachoeiras de Macacu) was performed in order to obtain income and anthropometric data, as well as fecal samples for parasitological analyses through the Ritchie technique., Results: Children infected with Ascaris lumbricoides had significantly lower means of height-for-age z scores (- 1.36 ± 0.75 vs. - 0.11 ± 1.02; p < 0.001), weight-for-age z scores (- 1.23 ± 0.74 vs. 0.09 ± 1.15; p = 0.001), and weight-for-height z scores (- 0.68 ± 0.44 vs. 0.23 ± 1.25; p = 0.006) when compared with uninfected children. Infection with hookworm was also associated with lower means of height-for-age z scores (- 1.08 ± 1.17 vs. - 0.12 ± 1.02; p = 0.015) and weight-for-age z scores (- 1.03 ± 1.13 vs. 0.08 ± 1.15; p = 0.012). Children infected with Entamoeba coli presented significantly lower means of height-for-age z scores (- 0.54 ± 1.02 vs. - 0.09 ± 1.02; p = 0.005) and weight-for-age z scores (- 0.44 ± 1.15 vs. 0.12 ± 1.15; p = 0.002). The multivariate multiple linear regression analysis showed that height-for-age z scores are independently influenced by monthly per capita family income (β = 0.145; p = 0.003), female gender (β = 0.117; p = 0.015), and infections with A. lumbricoides (β = - 0.141; p = 0.006) and Entamoeba coli (β = - 0.100; p = 0.043). Weight-for-age z scores are influenced by monthly per capita family income (β = 0.175; p < 0.001), female gender (β = 0.123; p = 0.010), and infections with A. lumbricoides (β = - 0.127; p = 0.012), and Entamoeba coli (β = - 0.101; p = 0.039). Monthly per capita family income (β = 0.102; p = 0.039) and female gender (β = 0.134; p = 0.007) positively influences mid upper arm circumpherence., Conclusions: Intestinal parasitism and low family income negatively influence the physical development of children in low-income communities in different Brazilian regions.

Background: Bovine leptospirosis is mainly characterized by reproductive disorders. Contamination of the oocyte was previously demonstrated in vitro, resulting in some apparent damage. However, it is not clear whether it occurs under natural conditions. The present study aimed to characterize the presence of pathogenic Leptospira DNA in the ovarian follicles of non-pregnant cows., Methods: Follicular fluid samples were collected from 65 animals and subjected to lipL32 PCR and secY sequencing., Results: In total, seven of 65 (10.8%) were positive, indicating a possible early infection of the oocyte. Moreover, secY sequencing identified L. interrogans and L. santarosai, both very closely related to bovine strains from the Sejroe serogroup (Hardjoprjitno and Guaricura). We demonstrated that ovarian follicles can also be infected., Conclusions: It was hypothesised that ovarian infection can contribute to embryonic death, causing reproductive failure and estrus repetition. In the present study, we show that the organism identified in the follicle is closely related to one that is known to be associated with reproductive disorders., (© 2021 British Veterinary Association.)

(1) Background: Dogs are known as hosts of Leptospira interrogans and can spread this bacterium to the environment. Although Canicola is responsible for determining chronic disease in dogs, when affected by incidental serogroups such as Icterohaemorrhagiae, acute disease may occur with a predominance of clinical signs with hepatic and renal changes. In endemic areas, it is a serious public health problem. Thus, this study aims to estimate the incidence and duration of elimination of leptospires in the urine of dogs, taking another step from a previous study of our group, by a longitudinal, long-term and molecular approach. (2) Methods: A total of 125 dogs without apparent symptoms of leptospirosis were included in the study. The dogs were all PCR-negative and seronegative at the beginning of the study. Blood samples were collected for hematological examinations and urine for amplification of the lipL32 gene by PCR at five different time points during one year. (3) Results: Out of the 125 dogs, 62 became lipL32 PCR-positive (48.8% (95% CI, 47.9-49.7%)) at some point during the study, distributed as follows: at day 0, all negative; day 90, 18/125 (14.4% (95% CI, 13.5-15.3%)); day 180, 18/125 (14.4% (95% CI, 13.5-15.3%)); day 270, 12/125 (9.6% (95% CI, 8.7-10.5%)); and day 365, 14/125 (11.2% (95% CI, 10.3-12.1%)). Out of the 62 amplicons, 22 were sequenced, targeting a short region of secY gene. Of these, 20 (90.9%) were identical to the L. interrogans serovar Icterohaemorrhagiae, while two (9.1%) were Leptospira noguchii . (4) Conclusions: The fact that the leptospires of the Icterohaemorrhagiae serogroup were characterized was unexpected, since the animals remained clinically asymptomatic during the study. The fact that asymptomatic dogs shed leptospires is not new, but the extent of this fact and the characterized strain is impressive, with an impact on public health that cannot be overlooked.

Background: Entamoeba species harbored by humans have different degrees of pathogenicity. The present study explores the intra- and interspecific diversity, phylogenetic relationships, prevalence and distribution of tetra- and octonucleated cyst-producing Entamoeba in different Brazilian regions., Methods: Cross-sectional studies were performed to collect fecal samples (n = 1728) and sociodemographic data in communities located in four Brazilian biomes: Atlantic Forest, Caatinga, Cerrado, and Amazon. Fecal samples were subjected to molecular analysis by partial small subunit ribosomal DNA sequencing (SSU rDNA) and phylogenetic analysis., Results: Light microscopy analysis revealed that tetranucleated cysts were found in all the studied biomes. The highest positivity rates were observed in the age group 6-10 years (23.21%). For octonucleated cysts, positivity rates ranged from 1 to 55.1%. Sixty SSU rDNA Entamoeba sequences were obtained, and four different species were identified: the octonucleated E. coli, and the tetranucleated E. histolytica, E. dispar, and E. hartmanni. Novel haplotypes (n = 32) were characterized; however, new ribosomal lineages were not identified. The Entamoeba coli ST1 subtype predominated in Atlantic Forest and Caatinga, and the ST2 subtype was predominant in the Amazon biome. E. histolytica was detected only in the Amazon biome. In phylogenetic trees, sequences were grouped in two groups, the first containing uni- and tetranucleated and the second containing uni- and octonucleated cyst-producing Entamoeba species. Molecular diversity indexes revealed a high interspecific diversity for tetra- and octonucleated Entamoeba spp. (H ± SD = 0.9625 ± 0.0126). The intraspecific diversity varied according to species or subtype: E. dispar and E. histolytica showed lower diversity than E. coli subtypes ST1 and ST2 and E. hartmanni., Conclusions: Tetra- and octonucleated cyst-producing Entamoeba are endemic in the studied communities; E. histolytica was found in a low proportion and only in the Amazon biome. With regard to E. coli, subtype ST2 was predominant in the Amazon biome. The molecular epidemiology of Entamoeba spp. is a field to be further explored and provides information with important implications for public health.

Soil-transmitted helminthiasis (STHs) are poverty-related diseases with high prevalence rates in developing countries. The present study aims to describe the epidemiological scenario of STHs in an urban population in the Brazilian Amazon. A cross-sectional survey ( n  = 349 children aged 1-15 years) was carried out to obtain faecal samples and sociodemographic and sanitation data. Among the children, 143 (41%) were positive for at least one STH. Prevalence rates of infections by A. lumbricoides , T. trichiura , and hookworms were 24.4%, 42.6%, and 9%, respectively. A logistic regression multivariate model showed that infection with A. lumbricoides is significantly more frequent in children aged 11-15 years (odds ratio [OR] = 2.38; 95% confidence interval [CI] = 1.15-4.94; p =0.018) and the presence of latrines inside houses is a protection factor against ascariasis (OR = 0.38; 95% CI = 0.17-0.85; p =0.019). Positivity for T. trichiura is higher in the 5-10 (OR = 3.31; 95% IC = 1.85-5.89; p =0.001) and 11-15 age groups (OR = 3.16; 95% IC = 1.66-6.00; p =0.001), in children living in poor families (OR = 1.78; 95% IC = 1.01-3.14; p =0.045) and practicing open evacuation (OR = 2.07; 95% IC = 1.07-3.99; p =0.029). Hookworm infection is more frequent in children aged 11-15 years (OR = 6.70; 95% IC = 1.91-23.43; p =0.002), males (OR = 6.35; 95% IC = 2.00-20.14; p =0.002), and those living in stilt houses (OR = 3.52; 95% IC = 1.22-10.12; p =0.019). The use of albendazole in the last six months was a protection factor against hookworm infection (OR = 0.31; 95% IC = 0.10-0.96; p =0.042). The proportion of mild, moderate, and severe infections was 55.2%, 37.8%, and 7%, respectively, for A. lumbricoides , 72.4%, 24.3%, and 3.3% for T. trichiura , and 93.8%, 3.1%, and 3.1% for hookworms. Significantly higher worm burdens in T. trichiura and hookworm infections were associated with practicing open defecation and living in stilt houses. The data points to the need to improve sanitation infrastructure in Amazonian cities with similar sociodemographic and environmental characteristics., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2021 Deiviane Aparecida Calegar et al.)

Leptospira is a bacteria that causes leptospirosis and is transmitted through water, soil, or mud that is contaminated by the urine of infected animals. Although it is mainly associated with the urban environment, Leptospires also circulate in rural and wild environments. This study aimed to investigate the role of small mammals in leptospirosis epidemiology in the western Amazon, Brazil. In total, 103 animals from 23 species belonging to the orders Didelphimorphia and Rodentia were captured. Blood, kidney, and urine samples were collected and Microscopic Agglutination Test (MAT), lip L32 PCR, sec Y sequencing, and culturing were conducted. MAT was reactive on 1/15 sera, and no bacterial isolate was obtained. PCR yielded 44.7% positive samples from 16 species. Twenty samples were genetically characterized and identified as L. interrogans ( n = 12), L. noguchii ( n = 4), and L. santarosai ( n = 4). No statistical association was found between the prevalence of infection by Leptospira spp. in small mammals within carrier/hosts species, orders, study area, and forest strata. Our results indicate a high prevalence of pathogenic Leptospira spp. in several rodent and marsupial species and report the first evidence of Leptospira spp. carrier/hosts in the Brazilian Western Amazon., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Medeiros, Braga Domingos, Azevedo, Peruquetti, de Albuquerque, D'Andrea, Botelho, Crisóstomo, Vieira, Martins, Teixeira, Carvalho-Costa and Lilenbaum.)

Background: In northeastern Brazil, the wild nature of Trypanosoma cruzi vectors has challenged control actions. This study aims to describe the entomological and epidemiological scenario of Chagas disease in rural communities in the state of Piauí., Methods: A cross-sectional study (n=683 individuals/244 dwellings) was carried out to obtain serum samples, sociodemographic data and intra- and peridomestic triatomines., Results: The overall seroprevalence rate was 8.1%, with no positive tests among subjects <30 y of age. Prevalence rates reached 34.3% and 39.1% among subjects 61-75 and >75 y of age, respectively; 1474 triatomines were collected, of which 90.3% were found in peridomiciliary structures and 9.7% inside houses; 87.2% were classified as Triatoma brasiliensis. T. cruzi infection rates in insects were 0.5% by light microscopy and 0.9% by culture in NNN/LIT medium. Five cultivated isolates were submitted to molecular genotyping, three of which were identified as T. cruzi I and two as T. cruzi II., Conclusions: Although no vector transmission currently occurs, prevalence rates are high in adults and the elderly. This disease should be targeted by primary healthcare providers. Insect surveillance and control activities should not be discontinued in an environment favourable to the perpetuation of house colonization by triatomines., (© The Author(s) 2020. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.)

Introduction: Papanicolaou test (Pap smear) is the standard screening test of pre-neoplastic lesions and cervical cancer. This study aimed to investigate cervical cancer screening results and risk factors such as age, reason for the examination, the epithelia detected in the sample, microbiota and signs of sexually transmitted infection (STIs) of women in a maternity school in Fortaleza, Ceará, Brazil., Methods: In this cross-sectional study, data were retrieved of 353 women who underwent Pap smear between April 2016 and January 2017 at the Federal University of Ceará., Results: Of all Pap smear samples retrieved, 54.1% (191/353) had glandular epithelium and 40.2% (142/353) had metaplastic epithelium. After statistical analyses adjusted for the final model, age ≥51 years (odds ratio = 3.47) and signs of STIs (odds ratio = 4.95) remained as risk factors., Conclusions: The diagnosis of high-grade lesions and carcinomas in patients older than 50 years indicates a deficiency in cervical screening. Women with signs and symptoms of STIs and candidiasis sought medical services more frequently than asymptomatic women, and presence of these signs and symptoms contributes to the diagnosis of cervical cancer. We highlight the importance of obtaining a correct smear sampling to allow prompt detection of all preneoplastic lesions; moreover, the implementation of human papillomavirus vaccination and an efficient routine Pap screening are necessary in low-resource settings., (© 2020 John Wiley & Sons Ltd.)

Bovine genital leptospirosis is a chronic disease that causes reproductive disorders such as abortions, stillbirths, and estrus repetition, as well as economic losses. Despite clinical signs related to reproductive failure, the majority of studies have focused on the detection of Leptospira spp. in the urine, while few have considered the reproductive tract. Consequently, the aim of the present study was to investigate the uterus as an important extra-renal site of leptospiral infection in cows. A total of 42 non-pregnant cows were studied at a slaughterhouse. Blood samples and uterine fragments were collected for serology and molecular analysis, respectively. Concerning serologic results, 20.5 % presented as reactive, all of them against the Sejroe serogroup. Regarding lipL32 PCR, 26.2 % (11/42) of samples were positive for pathogenic Leptospira sp. Sequencing the secY gene short region enabled nine strains to be characterized, all of which were L. interrogans, with high identity (98.8 %-99.8 %) with serovar Hardjo. The use of molecular tools substantially improved the sensitivity of Leptospira sp. detection at species level and demonstrated that the uterus is an important site of bovine leptospiral infection. The findings of the present study reinforce our understanding that leptospiral uterine infection are associated to members of the Sejroe serogroup., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Poverty, malnutrition and neglected tropical diseases such as soil-transmitted helminthiases (STHs) interact in a multi-causal feedback network. This study aimed to assess the relationships between STHs, income and nutritional status of children in impoverished communities in the city of Caxias, Maranhao State, Northeastern Brazil. A cross-sectional survey (n=259 children) was carried out with the collection of fecal samples and assessment of sociodemographic, anthropometric, dietary and sanitation data. Hookworm infection and ascariasis presented prevalence rates of 14.3% and 9.3%, respectively. The logistic regression analysis showed that hookworm infection was more frequent in males (odds ratio [OR]=3.43; 95% confidence interval [CI]=1.45-8.08), children aged 11-15 years old (OR=3.72; 95% CI=1.19-11.62), children living in poor families (OR=2.44; 95% CI=1.04-5.68) and those living in rented houses (OR=5.74; 95%CI=1.91-17.25). Concerning ascariasis, living in the Caldeiroes community (OR=0.01; 95%CI=0-0.17) and belonging to the 11-15 years age group (OR=0.21; 95%CI=0.04-1.02) were protection factors. Poor children have a significantly lower frequency of consumption of meat, milk, vegetables, tubers and fruits than not poor children. The frequent consumption of meat, milk and tubers was associated with significant higher values in the parameter height-for-age, whereas the consumption of meat and milk positively influenced the weight-for-age. The frequencies of stunting, underweight and wasting were 8.1%, 4.9% and 2.9%, respectively. The multivariate model demonstrated that stunting was significantly associated with economic poverty (OR=2.82; 95%CI=1.03-7.70) and low weight was associated with male sex (OR=6.43; 95% CI=1.35-30.68). In conclusion, the study describes the interactions between the dimensions of development represented by income, STHs and nutritional status revealing the importance of raising income levels to improve the living conditions of families in impoverished communities in Northeastern Brazil.

This study aims to assess the prevalence, distribution, and etiological profile of intestinal parasitism in children living in periurban areas in Cachoeiras de Macacu, Rio de Janeiro, Brazil. A community-based cross-sectional survey ( n  = 479) was carried out. Prevalence of infection with G. duodenalis and E. histolytica / E. dispar was 8.6% ( n  = 41) and 13.4% ( n  = 64), respectively. Infection with G. duodenalis was significantly more frequent among children living in poor families (24/187 (12.8%) vs. 16/272 (5.9%); prevalence ratio (PR) = 2.18; 95% confidence interval (CI) = 1.19-3.99; p =0.011). This difference was also significant for infection with any pathogenic parasite (43/187 (23%) vs. 40/272 (14/7%); PR = 1.56; 95% CI = 1.06-2.30; p =0.026). In addition, people residing in houses with more than four inhabitants showed significantly higher positivity for infections with G. duodenalis and with E. histolytica / E. dispar (22/138 (15.9%) vs. 16/311 (5.1%); PR = 3.09; 95% CI = 1.68-5.71; p < 0.001 for G. duodenalis and 32/138 (23.2%) vs. 30/311 (9.6%); PR = 2.40; 95% CI = 1.52-3.79; p < 0.001 for E. histolytica / E. dispar ). Laboratory diagnosis of protozoan enteric infections and effective drugs for their treatment are unmet goals in the primary health care system. Therefore, giardiasis and amebiasis are neglected conditions., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2020 Deiviane A. Calegar et al.)

Cryptococcosis, a potentially fatal mycosis in humans, is acquired via exposure to exogenous environmental sources. This study aimed to investigate the frequency, genetic diversity, and virulence of cryptococcal strains isolated from indoor dust in the Rio Negro micro-region of the Brazilian Amazon. A total of 8.9% of the studied houses were positive, recovering nine Cryptococcus neoformans VNI and 16 C. gattii VGII isolates, revealing an endemic pattern in domestic microenvironments. The International Society for Human and Animal Mycology (ISHAM) consensus multilocus sequence typing (MLST) scheme for the C.&nbsp;neoformans/C. gattii species complexes identified two sequence types (STs), ST93 and ST5, amongst C. neoformans isolates and six STs amongst C. gattii isolates, including the Vancouver Island Outbreak ST7 (VGIIa) and ST20 (VGIIb), the Australian ST5, and ST264, ST268 and ST445, being unique to the studied region. Virulence studies in the Galleria mellonella model showed that five C. gattii strains and one C. neoformans strain showed a similar pathogenic potential to the highly virulent Vancouver Island outbreak strain CDR265 (VGIIa). The findings of this study indicate that humans can be exposed to the agents of cryptococcosis via house dust, forming the basis for future studies to analyze the impact of early and continuous exposure to indoor dust on the development of subclinical or clinical infections., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript or in the decision to publish the results.

Leptospirosis presents a complex and dynamic epidemiology. Bovine leptospirosis has been described as a major infectious disease impairing reproductive efficiency. Although infections by Leptospira interrogans, L. santarosai and L. borgpetersenii are frequently reported in cattle, the presence of L. noguchii in these animals should not be neglected. In this study, we describe serological (MAT) and molecular characterization (rrs and secY gene sequencing, multilocus sequence typing [MLST] and pulsed-field gel electrophoresis [PFGE]) of eight L. noguchii strains obtained from slaughtered cows. Intraspecific genetic diversity was evaluated, and haplotype networks were constructed based on hosts and geographical localizations. Strains were characterized as belonging to serogroups Australis, Autumnalis and Panama, and molecular characterization showed a high heterogeneity of these strains. Ten different STs were found (including nine new STs and 39 novel alleles) as well as nine different pulsotypes. Two clonal complexes were found. Phylogenetic trees based on secY locus and concatenated MLST loci showed two main clusters, with sequences from the present study included in the first. In general, there was no relationship between the geographical origin and the secY phylogenetic clusters, as well as between secY phylogenetic clusters and serogroups. Molecular diversity indexes confirmed a high variability (H > 0.8). This high intraspecific variation observed may be related to differences in virulence, pathogenicity and antigenicity or even adaptability of the strains. In addition, haplotype networks clearly demonstrated the circulation of genotypes between humans and animals, confirming the zoonotic potential. The present study provides relevant data for the study of leptospirosis in the One Health context, where human, animal and environmental health is closely connected., (© 2019 Blackwell Verlag GmbH.)

This study aimed to describe the spatial distribution and assess entomological indicators of synanthropic triatomines in Piaui State, Northeastern Brazil. We used surveillance data on the detection, identification and assessment of natural infection with trypanosomatids from triatomines in the State from 2014 to 2017. The State was divided into four macroregions. In relation to the dispersion rates of triatomines, they were much lower in the North, when compared to Southwest, Southeast and Central North macroregions. Infestation rates were higher in the Southwest and Southeast and intradomicile infestation rates varied during the study period, reaching high values in all regions. Insects belonging to the species Triatoma brasiliensis complex, Triatoma pseudomaculata, Triatoma sordida, and to the genus Panstrongylus spp. and Rhodnius spp. were collected during this period. T. brasiliensis was collected from all four regions of the State, but more frequently in those located in the Southeast. A similar pattern was observed for T. pseudomaculata. T. sordida was detected in the municipalities in the Southeast and Southwest regions, and less frequently in the Central North municipalities. Rhodnius spp. was detected in the Central North and North regions, and Panstrongylus spp. in the Central North and Southeast regions. The highest trypanosomatid-positivity rate of T. brasiliensis and Panstrongylus spp. was in the Southeast region. A significant proportion of the municipalities of Piaui State presents entomological parameters that indicate a risk of Chagas disease by vector transmission.

Objectives: To describe aspects of the microcephaly epidemic in the state of Piauí., Methods: All cases of congenital microcephaly confirmed in the state between 2015 and 2016 were included (n=100). Investigation forms of the Regional Reference Center for Microcephaly were reviewed. Discarded cases (n=63) were used as a comparison group., Results: In October, November, and December 2015 incidence rates reached 4.46, 6.33 and 3.86/1000 live births, respectively; 44 cases were reported in the state capital. Among the mothers of confirmed and discarded cases, the frequency of skin rash during pregnancy was 50/97 (51.5%) and 8/51 (15.7%), respectively (p<0.001); 33 confirmed cases (35.9%) had a head circumference z-score between -2 and -3, 23 (25%) between -3 and -4, and 8 (8.7%) had a z-score of less than -4. Head computer tomography scans revealed calcifications in 78/95 (82.1%) cases. Lissencephaly, hydrocephalus and agenesis of the corpus callosum were also frequently observed. Ophthalmic findings included retinal pigment epithelium rarefaction and atrophy. Absence of otoacoustic emissions was observed in 21/70 cases. One newborn also presented lower limb muscle atrophy. There were no significant differences in vaccination rates for influenza, diphtheria-tetanus-acellular pertussis, and hepatitis B in either group., Conclusions: The state of Piauí, like others in the northeastern region, faced an epidemic of congenital microcephaly between 2015 and 2016, presumably related to congenital Zika virus infection, more intense in the capital. Current challenges include the improvement of vector control, basic research, scaling-up of diagnostic tools for pre-natal screening of Zika virus, vaccines, and health care for affected children., (Copyright © 2018. Published by Elsevier Editora Ltda.)

In this study, we assessed the genetic diversity of Ascaris lumbricoides / Ascaris suum circulating in humans and pigs, exploring potential zoonotic cycles in endemic areas in Brazil. We carried out cross-sectional surveys in four municipalities: Santa Isabel do Rio Negro (SIRN-AM) (n = 328); Nossa Senhora de Nazaré (NSN-PI) and Teresina (TER-PI) (n = 605 and n = 297, respectively); and Cachoeiras de Macacu (CAM-RJ) (n = 543). We also studied 61 fecal samples/adult worms obtained from pigs (n = 53 in NSN-PI and n = 8 in TER-PI). A ~450 bp fragment of the Ascaris cytochrome c oxidase subunit 1 (cox1) and ~400 bp of the NADH dehydrogenase subunit 1 (nad1) were amplified and sequenced. Maximum-likelihood (ML) tree and Median-joining (MJ) haplotype network analyses were performed. We also performed scanning electron micrographs of adult specimens. Positivity rates were 93/328 (28.4%) in SIRN-AM, 6/297 (2.0%) in TER-PI, 0/605 (0%) in NSN-PI, and 6/543 (1.1%) in CAM-RJ. In NSN-PI it reached 11/53 (20.7%) in pigs. The MJ network based on cox1 locus (383 bp) revealed three main clusters, one centered around haplotypes H01/H28/H32 and the other around H07/H11. The cox1 haplotypes had a heterogeneous distribution, showing no pattern by geographic region, and high haplotype diversity. The ML trees based on cox1 and nad1 loci showed a similar topology with each other, and with the haplotype networks. Three distinct clusters were observed. Sequences of cox1 and nad1 from humans and animals were distributed throughout the tree and it was not possible to differentiate specimens of human and swine origin. Ascaris populations obtained from humans and swine in different Brazilian regions are not discriminable through the genetic markers used, which indicates the potential for zoonotic transmission and the need for better control of these infections in swine herds, mainly when created in a peridomestic environment., Competing Interests: The authors have declared that no competing interests exist.

Leptospirosis is a common worldwide bacterial zoonosis and has been studied in One Health approaches. Small mammals are described as the most important maintenance reservoirs of several pathogens in nature, including leptospires. The aim of this study was to identify infection by leptospires among small mammals on the Atlantic forest biome and evaluate their potential as carriers of these spirochetes. A total of 153 small mammals belonging to orders Rodentia and Didelphimorphia (distributed on 17 genera and 22 species) were captured. Blood and kidney samples were collected from animals and a conventional PCR targeted on lipL32 gene was conducted on renal tissues. Species identification was performed in eight samples by sequencing of rrs gene. A total of 28% of the animals presented lipL32 PCR-positive, and four pathogenic Leptospira species (L. interrogans, L. borgpetersenii, L. santarosai and L. noguchii) were identified. This study highlights the role of small mammals as carriers of leptospires on the Atlantic Forest representing a potential source of pathogenic Leptospira spp infection for both humans and domestic animals., (© 2019 Blackwell Verlag GmbH.)

Hookworm infection persists focally in rural communities in Brazil. In this study, we analyze the mitochondrial nucleotide sequences obtained from hookworms infecting humans in order to characterize species composition and assess their genetic diversity and phylogenetic relationships. Field expeditions and cross-sectional surveys were carried out in three Brazilian municipalities from 2013 to 2017: Nossa Senhora de Nazaré (n = 605) and Teresina (n = 297), in the state of Piauí, and Russas (n = 213) in the State of Ceará. Parasitological methods were used to evaluate fecal samples. Hookworm-positive samples had a partial mtDNA cox1 amplified and sequenced. Maximum-likelihood and Bayesian analysis demonstrated two strongly-supported clades, including Group A, corresponding to Necator americanus, and Groups B and C, corresponding to Necator sp. Group A was divided into three main clusters: A1 grouped with Asian sequences, A2 grouped with African sequences, and A3 had only Asian sequences. Group B was closely related to Necator sp., showing a sequence similarity of 98%-99% with African samples circulating zoonotically among humans and non-human primates. Twenty three N. americanus haplotypes were identified. N. americanus Median-Joining network revealed three distinct groups, designated again as A1, A2, and A3. Group A1 presented a star-like shape, with one dominant haplotype. The molecular dating suggested that the two clades dividing N. americanus and Necator sp. began to diverge during the middle Pleistocene. The most recent common ancestor among N. americanus groups was dated to the late Pleistocene. Hookworms circulating in the studied communities are structured in well-defined subpopulations presenting both Asian and African genetic backgrounds. This reveals a double origin for hookworms in northeastern Brazil and opens up new possibilities in phylogeographic, evolutionary, and molecular epidemiological studies in regions where hookworms persists focally, despite control efforts. The presence of potentially zoonotic species and the specific identification of Necator sp. should be further investigated., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Chagas disease is one of the most significant systemic parasitosis in Latin America, caused by Trypanosoma cruzi , which is mainly transmitted by hematophagous insects, the triatomines. This research was carried out in both domestic and wild environments throughout a Northeastern rural locality. Triatomines were captured in both peridomicile and wild environments, obtaining 508 specimens of triatomines, of which 99.6% were Triatoma brasiliensis . Insects were captured in 10 (18.5%) peridomiciles with an average of 8.3 triatomines per residence. Triatoma brasiliensis nymphs and adults were found in six peridomiciles, generating a 11.1% colonization. No T. cruzi infection was detected in the 447 peridomestic insects analyzed. On the other hand, of the 55 sylvatic T. brasiliensis molecularly examined for T. cruzi , 12 (21%) were positive, all harboring T. cruzi I. The blood meal analysis by enzyme-linked immunosorbent assay from gut content revealed that both peridomestic and wild triatomine populations fed mainly on birds, refractory to the parasite, which may explain the null rate of natural infection prevalence in the domestic environment. However, infected triatomines for potential home infestation within the radius of insect dispersion capacity were registered in rock outcrops around the dwellings. Anthropogenic environmental influences are able to rapidly alter these scenarios. Therefore, to avoid disease transmission to humans, we recommend constant vector control combined with periodic serological surveillance. The associated methodology presented herein may serve as a model for early detections of risk factors for Chagas disease transmission in the Brazilian Northeast.