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Background: Synthetic peptides of tandem repeat proteins (TRPs) have been employed in the serologic analysis of canine monocytic ehrlichiosis (CME) and used in epidemiological studies in Brazil. Based on molecular studies of TRPs, different genotypes of Ehrlichia canis have been described, but data on their pathogenicity remain unknown. Objectives: To correlate hepatic, renal, and muscular alterations in relation to different genotypes of E. canis in naturally exposed dogs using enzyme‐linked immunosorbent assay (ELISA) with TRP19 and TRP36 synthetic protein antigens. Methods: Two hundred serum samples were subjected to ELISA with the antigens of TRP19 and three genotypes (US, Br, and CR) of TRP36 of E. canis circulating in Brazil. Positive sera were evaluated through eight biochemical parameters, and the results were evaluated by principal component analysis and canonical correlation. Results: ELISA revealed that 47 (23.5%) serum samples reacted to the BrTRP36 peptide, 36 (18%) reacted to the TRP19 peptide, and 8 (4%) reacted to the USTRP36 and CRTRP36 peptides separately. The most frequent biochemical alterations observed were for CK (59.4%), ALB (31.8%), GLO (28.9%), TP (28.9%), ALP (26%), urea (24.6%), creatinine (14.4%), and ALT (14.4%). The most prominent diagnostic method in canonical correlation analysis was BrTRP36, followed by TRP19, which correlated with hyperglobulinemia and hypoalbuminemia. Conclusions: Antibodies that reacted against the Brazilian genotype of E. canis correlated positively with hyperglobulinemia and increases in serum urea and creatinine. According to our results, the Brazilian genotype of E. canis is related to the chronic phase of CME. [ABSTRACT FROM AUTHOR]

The aim of the present study was to characterize (phenotypically and genotypically) two strains of Brucella abortus identified as belonging to biovar 4 isolated from cattle in Brazil. The strains were isolated from cervical bursitis from cattle in the states of Pará and Rio Grande do Sul, respectively. In the phenotypic identification, the isolates were positive in CO2 requirement, produced H2S, were resistant to basic fuchsin (20 µg / mL) and sensitive to thionin (20 µg / mL and 40 µg / mL) and presented M surface antigen, but A surface antigen is absent. The isolates were positive in the PCR for the bcsp31 gene (genus-specific) and in the AMOS-enhanced PCR, both isolates showed a band profile consistent with B. abortus biovar 1, 2 or 4. Moreover, both isolates also showed restriction patterns identical to the reference strain when tested by the omp2b PCR-RFLP. In genotyping using Multiple Locus Variable Number of Tandem Repeat (VNTR) Analysis - MLVA (MLVA16), the isolates showed differences in several loci (Bruce42, Bruce19, Bruce04, Bruce16 and Bruce30); by Multiple Locus Sequence Typing (MLST), they also exhibited differences in sequence type (ST), strain 16/02 ST1 (2-1-1-2-1-3-1-1-1) and strain 128/11 ST (22-1-1 -8-9-3-1-1-1). The extensive typing of B. abortus strains isolated from cattle in Brazil using different approaches confirmed the occurrence of rare B. abortus biovar 4 in the country. [ABSTRACT FROM AUTHOR]

Erwinia psidii is the causal agent of bacterial blight of guava, and wilt and dieback of eucalypt in Brazil. Previous studies using repetitive sequence‐based PCR (rep‐PCR) revealed limited genetic diversity in E. psidii populations. Here, the draft genome sequence of the type strain IBSBF435 was probed for variable number of tandem repeats (VNTRs) that could be used to study E. psidii genetic diversity by multilocus variable number of tandem repeat analysis (MLVA). Eleven selected VNTRs were used to assess genetic relatedness among 108 E. psidii strains from guava (36) and eucalypt (72), of different geographic origins and years of collection. A total of 79 haplotypes were detected in the strain collection. The number of alleles per locus ranged from 3 to 15. A minimum spanning tree indicated the occurrence of nine clonal complexes, of which two contained only guava strains, five only eucalypt strains, and two contained guava and eucalypt strains. The three oldest strains collected from guava in the 1980s (IBSBF435, IBSBF454, and IBSBF493) exhibited distinct haplotypes. IBSBF435 was genetically more closely related to some eucalypt strains than to guava strains, which supports previously reported lack of host specificity. IBSBF454 grouped with guava strains more recently collected from different states, suggesting that it could have been disseminated to different Brazilian areas. The implication of the lack of differentiation between guava and eucalypt strains is discussed in relation to the detection of E. psidii affecting eucalypt more than 25 years after the first description of guava bacterial blight. [ABSTRACT FROM AUTHOR]

Life-threatening bacterial infections are a major concern in health care services worldwide. This retrospective study aimed to demonstrate genetic and biochemical diversity in isolates of Acinetobacter baumannii and Pseudomonas aeruginosa from a public hospital in Brazil. A total of 63 isolates collected from different sites of infection and hospital sectors were characterized, and their susceptibility profile to antibiotics was assessed for 18 drugs belonging to 8 antimicrobial categories using the automated BACTEC system. Genetic diversity was assessed using the multiple locus variable number tandem repeat analysis. Among the isolates of A. baumannii, 83% were classified as extensively drug resistant (XDR), and 17 genotypic profiles were identified. About 67% of P. aeruginosa isolates were susceptible to antimicrobials and were distributed into 37 genotypic profiles, revealing genetic heterogeneity. This study has demonstrated the multicolonization of investigated pathogens and the high frequency (95.8%) of multidrug-resistant and XDR, as well as high genetic diversity, among the isolates supporting the continuous need to monitor these species in the hospital environment. [ABSTRACT FROM AUTHOR]

• First report of tick-borne pathogens in carthorses from a tri-border area of Brazil. • Overall, 54.42% carthorses were positive for at least one tick-borne pathogen. • Dermacentor nitens ticks were found parasitizing carthorses. • The low Ehrlichia spp. seroprevalence may be due to the absence of Amblyomma ticks. • Hemoplasma infection in horses may be considered accidental or an uncommon infection. Tick-borne diseases (TBD) constitute an important group of illness affecting animals and humans worldwide. In Brazil, carthorses are frequently exposed to ticks and tick-borne pathogens, leading to impairment of horse performance and imposing restrictions by the international veterinary authorities for the importation of horses. Accordingly, this study has aimed to i) determine the prevalence of the TBD agents Theileria equi , Babesia caballi , Ehrlichia spp., and hemotropic mycoplasmas in carthorses, ii) identify the tick species parasitizing the animals, and iii) determine factors associated with exposure/infection in Foz do Iguaçu City, Parana state, southern Brazil. A total of 103 carthorses were screened for anti- T. equi and anti- Ehrlichia spp. antibodies by indirect fluorescent antibody assays (IFA). Samples were also tested by PCR assays targeting the 18S rRNA gene of T. equi and B. caballi , and 16S rRNA gene of hemoplasmas. Additionally, PCR assays targeting the 16S rRNA, disulfide bond formation protein (dsb) and tandem repeat proteins 36 (trp36) genes of Ehrlichia spp. were also performed. Antibodies to T. equi and Ehrlichia spp. were detected in 43/103 (41.75%; 95% CI: 32.10–51.88%) and 5/103 (4.85%; 95% CI: 1.59–10.97%) horses by IFA, respectively. DNA of T. equi and B. caballi were found in 25/103 (24.27%; 95% CI: 16.36–33.71%) and 10/103 (9.71%; 95% CI: 4.75–17.13%) carthorses, respectively, and all tested negative for Ehrlichia spp. and hemoplasmas. All sequences showed ≥99% identity with multiple T. equi and B. caballi 18S rRNA gene sequences deposited in GenBank. Overall, 191 Dermacentor nitens ticks were collected from 25/103 (24.27%) animals. Carthorses older than 5 years were more likely to be positive for T. equi (p < 0.05). In conclusion, equine piroplasmosis agents are highly prevalent in carthorses from Foz do Iguaçu City. The low prevalence of Ehrlichia spp. found may be due to the absence of Amblyomma ticks infesting animals, which should be further investigated. [ABSTRACT FROM AUTHOR]

The pathovar viticola of Xanthomonas citri causes bacterial canker of grapevine. This disease was first recorded in India in 1972, and later in Brazil in 1998, where its distribution is currently restricted to the northeastern region. A multilocus sequence analysis (MLSA) based on seven housekeeping genes and a multilocus variable number of tandem repeat analysis (MLVA) with eight loci were performed in order to assess the genetic relatedness among strains from India and Brazil. Strains isolated in India from three related pathovars affecting Vitaceae species and pathogenic strains isolated from Amaranthus sp. found in bacterial canker-infected vineyards in Brazil were also included. MLSA revealed lack of diversity in all seven genes and grouped grapevine and Amaranthus strains in a monophyletic group in X. citri. The VNTR (variable number of tandem repeat) typing scheme conducted on 107 strains detected 101 haplotypes. The total number of alleles per locus ranged from 5 to 12. A minimum spanning tree (MST) showed that Brazilian strains were clearly separated from Indian strains, which showed unique alleles at three loci. The two strains isolated from symptomatic Amaranthus sp. presented unique alleles at two loci. STRUCTURE analyses revealed three groups congruent with MST and a fourth group with strains from India and Brazil. Admixture among populations were observed in all groups. MST, STRUCTURE and e-BURST analyses showed that the strains collected in 1998 belong to two distinct groups, with predicted founder genotypes from two different vineyards in the same region. This suggest that one introduction of grape planting materials contaminated with genetically distinct strains took place, which was followed by pathogen adaptation. Genome sequencing of one Brazilian strain confirmed typical attributes of pathogenic xanthomonads and allowed the design of a complementary VNTR typing scheme dedicated to X. citri pv. viticola that will allow further epidemiological survey of this genetically monomorphic pathovar. [ABSTRACT FROM AUTHOR]

Abstract: Canine monocytic ehrlichiosis is a highly prevalent disease in Brazil, where the genetic diversity of Ehrlichia canis remains undefined. In this study, we used the TRP36 gene to examine the genetic diversity of E. canis strains from naturally infected dogs residing in five distinct geographic regions in Brazil. E. canis DNA was detected in 82/126 (65%) dogs by dsb-specific PCR and E. canis was isolated in cell culture from 13 dogs. Sequences obtained from dsb genes amplified from the isolates were identical to the US E. canis strain. An extended molecular characterization based on the TRP36 gene identified two major genogroups based on differences among eight isolates. Isolates with tandem repeat amino acid sequence (TEDSVSAPA) identical to the previously reported TRP36 sequence were found in the midwest, northeast and southeast regions of Brazil, and classified into the US genogroup. A novel Brazilian genotype with a different tandem repeat sequence (ASVVPEAE) was also identified in midwest, northern and southern regions. Similarity in the N-terminal sequence of a US genogroup member with the Brazilian genogroup suggested that genomic recombination between the two genogroups may have occurred. Other subtypes within the Brazilian genogroup were also identified using C-terminal amino acid divergence. We identified two distinct major Brazilian genogroups and several subtypes based on analysis of TRP36, and such information will be useful for further genotyping and possible associations with disease severity, understanding of the genetic and antigenic variability of E. canis, and for developing strain-specific vaccines and diagnostic methods based on TRP36. [Copyright &y& Elsevier]

Abstract: Salmonella Enteritidis (S. Enteritidis) is a major causative agent of food-borne gastroenteritis associated with the consumption of contaminated poultry products. In this study we used multilocus variable number of tandem repeats (VNTRs) analysis (MLVA) to discriminate a total of 188 S. Enteritidis strains recovered from human (n=67), food (n=61) and chickens (n=60) during a 24year period (1986 through 2010) in Brazil. MLVA profiles of the 188 strains from Brazil were compared to the MLVA profiles of 100 human clinical (n=52) and poultry-associated (n=48) strains isolated in North America between 1986 and 2008. MLVA typing led to classification of the 288 strains from Brazil and North America into two major clusters named A and B with 35% of similarity. Cluster A consisted of a vast majority of strains isolated from North America (n=71) and only three strains isolated from Brazil which included two pre-pandemic strains (SE5 and SE4). In contrast, cluster B consisted of all of the post-pandemic strains isolated from Brazil (n=185) and fewer strains isolated from North America (n=29). In general, MLVA typing showed that the North American strains were more genetically diverse whereas Brazilian strains were more genetically clonal. The clustering of pre-pandemic strains from Brazil with the North American strains suggests the possibility that the pre-pandemic strains were more likely genetically diverse; however after 1993 a new and prevalent subtype of S. Enteritidis was introduced in this country. This is the first study describing MLVA genotyping of the S. Enteritidis strains isolated from Brazil. [Copyright &y& Elsevier]

Background: A PCR-enzyme-linked immunosorbent assay (PCR-ELISA) was developed to overcome the need for sensitive techniques for the efficient diagnosis of Schistosoma infection in endemic settings with low parasitic burden. Methodology/Principal Findings: This system amplifies a 121-base pair tandem repeat DNA sequence, immobilizes the resultant 5′ biotinylated product on streptavidin-coated strip-well microplates and uses anti-fluorescein antibodies conjugated to horseradish peroxidase to detect the hybridized fluorescein-labeled oligonucleotide probe. The detection limit of the Schistosoma PCR-ELISA system was determined to be 1.3 fg of S. mansoni genomic DNA (less than the amount found in a single cell) and estimated to be 0.15 S. mansoni eggs per gram of feces (fractions of an egg). The system showed good precision and genus specificity since the DNA target was found in seven Schistosoma DNA samples: S. mansoni, S. haematobium, S. bovis, S. intercalatum, S. japonicum, S. magrebowiei and S. rhodaini. By evaluating 206 patients living in an endemic area in Brazil, the prevalence of S. mansoni infection was determined to be 18% by examining 12 Kato-Katz slides (41.7 mg/smear, 500 mg total) of a single fecal sample from each person, while the Schistosoma PCR-ELISA identified a 30% rate of infection using 500-mg of the same fecal sample. When considering the Kato-Katz method as the reference test, artificial sensitivity and specificity rates of the PCR-ELISA system were 97.4% and 85.1%, respectively. The potential for estimating parasitic load by DNA detection in feces was assessed by comparing absorbance values and eggs per gram of feces, with a Spearman correlation coefficient of 0.700 (P<0.0001). Conclusions/Significance: This study reports the development and field evaluation of a sensitive Schistosoma PCR-ELISA, a system that may serve as an alternative for diagnosing Schistosoma infection. Author Summary: Schistosomiasis is a neglected disease caused by worms of the genus Schistosoma. The transmission cycle requires contamination of bodies of water by parasite eggs present in excreta, specific snails as intermediate hosts and human contact with water. Fortunately, relatively safe and easily administrable drugs are available and, as the outcome of repeated treatment, a reduction of severe clinical forms and a decrease in the number of infected persons has been reported in endemic areas. The routine method for diagnosis is the microscopic examination but it fails when there are few eggs in the feces, as usually occurs in treated but noncured persons or in areas with low levels of transmission. This study reports the development of the PCR-ELISA system for the detection of Schistosoma DNA in human feces as an alternative approach to diagnose light infections. The system permits the enzymatic amplification of a specific region of the DNA from minute amounts of parasite material. Using the proposed PCR-ELISA approach for the diagnosis of a population in an endemic area in Brazil, 30% were found to be infected, as compared with the 18% found by microscopic fecal examination. Although the technique requires a complex laboratory infrastructure and specific funding it may be used by control programs targeting the elimination of schistosomiasis. [ABSTRACT FROM AUTHOR]

Anaplasma marginale is an obligate intracellular Gram-negative bacterium that is parasitic to erythrocytes and is the main agent of bovine anaplasmosis. This disease causes severe anemia and reduces weight gain and milk production, thus giving rise to major economic losses relating to livestock worldwide. The genetic diversity of this bacterium has been characterized based on sequences of major surface proteins (MSPs), especially MSP1α. This has enabled identification of several geographical strains, according to different amino acid sequences. The aim of this study was to investigate the genetic diversity of A. marginale in naturally infected Angus beef cattle during a disease outbreak in southeastern Brazil. Four blood samples were collected over a four-month period from each of 20 animals on a cattle farm in Itú, São Paulo, Brazil. Serum samples were subjected to indirect ELISA to detect anti-A. marginale IgG antibodies. The 80 whole-blood samples obtained were subjected to DNA extraction, quantitative real-time PCR (qPCR) for the msp1β gene, semi-nested PCR (snPCR) for the msp1α gene, cloning of the target fragment and sequencing using the Sanger method. The sequences obtained were analyzed for genetic diversity using the RepeatAnalyzer software. Both iELISA tests, using recombinant MSP5 and the Anaplasma antibody test kit (VMRD), revealed high seroprevalence: 91.25% and 97.5%, respectively. In qPCR, 100% of the samples were positive, with between 10 3 and 10 7 DNA copies/μL. In the snPCR based on the msp1α gene, 57.5% (46/80) of the samples were positive. Microsatellite analysis on the 36 sequences obtained showed the presence of genotypes H (58.3%), F (25%), E (19.4%), C (2.7%) and G (2.7%). The RepeatAnalyzer software identified 36 strains in the study region, among which some had not previously been described in the literature (13 27 13 27 13 F; 16 FF; τ 27; 63 29 104 29; LJ1 13 LJ1 13; 16 F 17; 16 F 91). High genetic diversity of A. marginale bacteria was found on this farm in Itú, São Paulo., (Copyright © 2021. Published by Elsevier GmbH.)

• A Leptospira strain was recovered from urine of a water buffalo. • The isolate was assigned to the species L. borgpetersenii and the Sejroe serogroup. • This was the first isolation of a strain other than the subgroup Wolffi in Brazil. • The strain was detected in the reproductive tract of experimentally infected hamsters. The isolation of leptospires from buffaloes worldwide is still limited to a few strains. Thus, the aim of this study was to describe the first Leptospira isolate from buffalo urine, assigned to the Sejroe serogroup, which does not belong to the Wolffi subgroup, traditionally isolated in Brazil. A total of 244 urine samples of water buffaloes (Bubalus bubalis) raised in the Brazilian Amazon were subjected to bacteriological culturing and polymerase chain reaction (PCR) for the detection of leptospires. The obtained isolate was characterized by serogrouping using polyclonal antibodies, partial DNA sequencing, Hardjo-Bovis-specific PCR, multiple-locus variable-number tandem repeat analysis (MLVA/VNTR) and experimental infection in hamsters. PCR was performed on the urine samples; 11/244 were positive (4.5 %) for Leptospira , and only one isolate was recovered (0.4 %). Regarding characterization, the isolate was assigned to the Sejroe serogroup with high titers (12,800) for the Saxkoebing and Sejroe serovar antisera. The isolate was negative for Hardjo-Bovis-specific PCR, and the species Leptospira borgpetersenii was identified by DNA sequencing. The MLVA results showed that the VNTR profile of the isolate was 1−2-5, compatible with that of serovars Sejroe/Istrica. In the experimental infection in hamsters, the animals did not develop clinical signs, and no macroscopic lesions were observed on the organs at necropsy; however, the strain was detected in the kidneys, uterus, and testicles of the animals. The isolate described herein highlights infection by Sejroe strains that may be overlooked in buffaloes and that may be different from those normally isolated and used in serological studies. [ABSTRACT FROM AUTHOR]

• 52 Leptospira isolates recovered from cattle raised in the Brazilian Amazon were characterized. • The isolates belonged to different species and serogroups. • Strains from the Sejroe serogroup were isolated for the first time in Brazilian Amazon. • Some genotypes may represent new strains not yet characterized. In Brazil, there have been few leptospires isolated from cattle, especially in the Amazon, implying that the epidemiology of the disease in this region is still largely unclear. In a previous study, 52 Leptospira isolates were obtained from urine of cattle raised in the Brazilian Amazon and, to achieve a greater understanding of Leptospira infection in cattle of this region, the present study aimed to serologically and molecularly characterizes all these isolates. The laboratory assays used were the microscopic agglutination test (MAT) adopting a panel of polyclonal antisera against Leptospira spp. for serogrouping the isolates, DNA sequencing (secY) and multiple locus variable number tandem repeat analysis (MLVA). The isolates belonged to five species: 20/52 were identified as L. borgpetersenii (38.5 %); 18/52 as L. kirschneri (34.6 %); 9/52 as L. santarosai (17.3 %); 3/52 as L. noguchii (5.8 %) and 2/52 as L. interrogans (3.8 %). With serogrouping, nine different serogroups were detected, with a high frequency of the Sejroe serogroup. MLVA showed that all L. borgpetersenii isolates had a profile compatible with serovar Hardjo; moreover, the other isolates demonstrated a diversity of patterns, and some of them may represent strains not yet characterized. In the Brazilian Amazon, the leptospires circulating in cattle revealed the unique aspects of infections in this area which, in addition to a variety of strains, were characterized by a high frequency of the Sejroe serogroup, highlighting the serovar Hardjo, which has not been reported in other regions of Brazil. [ABSTRACT FROM AUTHOR]

Short tandem repeat (STR) polymorphisms have been extensively used in forensic genetics analysis. Knowledge about the locus-specific mutation rates of STRs improves forensic probability calculations and interpretations of diversity data. To incorporate single-locus diversity information into autosomal STR mutation rate estimations, 13 STR loci were studied during 2007-2009 in 10,959 paternity investigation cases from Rio Grande do Sul, the southernmost state of Brazil, covering an overall number of 284,934 allelic transfers. A total of 355 mutations were identified; 348 repeats were gains or losses of one step, three were gains or losses of two steps, and four were gains or losses of not stepwise mutation. The mutation rates ranged from 4.6 × 10 to 2.3 × 10, and the overall mutation rate estimate was 1.2 × 10. The average of the paternal mutation rate (1.8 × 10) was five times higher than the maternal rate (0.36 × 10). The observed mutational features for STRs have important consequences for forensic applications, including the definition of criteria for exclusion in paternity testing and the interpretation of DNA profiles in identification analysis. [ABSTRACT FROM AUTHOR]

• Analysis of MLVA-16 revealed a high diversity among B. ovis isolates. • Argentinean isolates are related to B. ovis from New Zealand and Australia. • Isolates from Argentina, Uruguay and Brazil were grouped into two clades. • This work constitutes the largest study of B. ovis molecular genotyping until now. Brucellosis in rams is caused by Brucella ovis or Brucella melitensis and it is considered one of the most important infectious diseases of males in sheep-raising countries. Molecular characterization of Brucella spp. achieved by multi-locus variable number of tandem repeats analysis (MLVA) is a powerful tool to genotype Brucella spp. However, data regarding B. ovis genotyping is scarce. Thus, the aim of this study was to characterize the molecular diversity of B. ovis field-strains in Argentina. A total of 115 isolates of B. ovis from Argentina and Uruguay were genotyped using MLVA-16 and analyzed altogether with 14 publicly available B. ovis genotypes from Brazil. The Discriminatory Power (D) was 0.996 for MLVA-16 and 0.0998 for MLVA-8 and MLVA-11. Analysis of MLVA-16 revealed 100 different genotypes, all of them novel, including 90 unique ones. There was no correlation between geographical distribution and genotype and results showed a higher diversity within provinces than between provinces. Clustering analysis of the strains from Argentina, Uruguay and Brazil revealed that the 129 isolates were grouped into two clades. Whole Genome Sequencing analysis of the 19 B. ovis genomes available in public databases, and including some of the Argentinian strains used in this study, revealed clustering of the Argentinian isolates and closer relationship with B. ovis from New Zealand and Australia. This work adds new data to the poorly understood distribution map of genotypes regionally and worldwide for B. ovis and it constitutes the largest study of B. ovis molecular genotyping until now. [ABSTRACT FROM AUTHOR]

Coxiella burnetii, the zoonotic agent of Q fever, has a worldwide distribution. Despite the vast information about the circulating genotypes in Europe and North America, there is a lack of data regarding C. burnetii strains in South America. Here, we show the presence of novel multispacer sequence typing (MST) genotypes of C. burnetii in two clusters detected in Brazil and Argentina that seem to be distant in parenthood. Argentinian strains isolated from a tick belongs to a new phylogenetic branch of C. burnetii, and the Brazilians strains may be related to MST 20 and 61. Multilocus variable number tandem repeats analysis (MLVA) typing provided a deeper resolution that may be related to host clusters of bovines, caprine, ovine, and ticks. Our results corroborate with the reports of geotypes of C. burnetii. Thus, we highlight the need for more genotyping studies to understand the genetic diversity of C. burnetii in South America and to confirm the hypothesis of host-related genotypes. We also emphasize the importance of virulence studies for a better understanding of Q fever in the region, which may help in surveillance and disease prevention programs. [ABSTRACT FROM AUTHOR]

There is a significant interest in the contribution of the circadian timing system to individual differences in a wider range of behaviour. This is in part because sleep complaints and disturbances of the circadian timing system are common in psychiatric disorders. Based on diurnal preference, people can be divided into chronotypes with demonstrated differences in sleep-wake patterns and a variety of circadian rhythms, behavioral rhythms, and diurnal variation in mood. Specifically, greater eveningness has been related to greater depression. The human PER3 gene contains a variable number tandem repeat (VNTR) which is associated with diurnal preference: the longer allele (five tandem repeats) has been reported to associate with greater morningness, and the shorter allele (four tandem repeats) with greater eveningness. We have investigated the PER3 polymorphism and its relationship to sleep, circadian and depression characteristics in a Brazilian family-based cohort. Baependi is a small rural town in Brazil that provides a window of opportunity to study the influence of sleep circadian patterns in a highly admixed rural population with a conservative lifestyle. Here, we studied 786 subjects (mean age±SD 46.5±16.2, 42% female). Blood samples were collected from all participants. The Per3 length polymorphism was genotyped using PCR followed by gel electrophoresis. We tested for associations between the PER3 polymorphism and sleep characteristics, daytime sleepiness, and chronotype using the Pittsburgh Sleep Quality Index, Epworth Sleepiness Scale and Morningness-Eveningness Questionnaire, respectively. Depressive symptoms were assessed using the Beck Depression Inventory [BDI-II]). Our genotyping data showed that 36% of the subjects were homozygous for the shorter allele (4/4), 51% of the participants were heterozygous (4/5) and 12% of the subjects were homozygous for the longer allele (5/5). Allele frequency was 0.62 for the 4-repeat and a 0.38 for the 5-repeat. The percentages of the chronotypes Morning type (M-type), Neither type (N-type) and Evening type (E-type) were 5%, 46% and 7%, respectively. We observed an increase of the depression symptoms in evening types [F(2, 1014)=4.8, p=0.007]. On the other hand, no such associations were observed the PER3 genotype. There were no significant differences in the MEQ score [F(2, 317)=0.46, p=0.63], daytime sleepiness [(F(2, 317)=0.23, p=0.79)], sleep efficiency [(F(2, 317)=2.59, p=0.07)] and sleep characteristics [(F(2, 317)=1.92, p=0.14)] between the PER3 genotypes. We observed an association between eveningness and depression. Identifying factors contributing to individual differences in sleep and to describe how these factors are associated with physiological and psychological profiles may aid our understanding of the role of sleep and the circadian system in mental health. On the other hand, we were not able to replicate the association between the PER3 VNTR polymorphism and chronotype, which has been confirmed independently in multiple populations worldwide (including in Brazil). Neither did we observe any association with depression. CNPq – PVE 400791/2014-5. [ABSTRACT FROM AUTHOR]

There are few studies on the genetic diversity of Anaplasma marginale in Brazilian cattle herds, especially about beef cattle. The objective of this study was to evaluate the genetic diversity of A. marginale, based on the msp1α gene in Bos taurus indicus sampled from the Brazilian Pantanal. Aliquots of blood with and without EDTA were taken from 400 cattle (200 cows and 200 calves) across five extensive farms. The samples were submitted to the indirect immunoenzymatic assay (iELISA), quantitative real-time PCR (qPCR) for the msp1β gene and to the semi-nested (sn) PCR for the msp1α gene. Positive samples were sequenced by the Sanger method and subjected to diversity analysis using the RepeatAnalyser software. The percentage of positive animals by iELISA, qPCR and (sn) PCR was 72.2% (289/400), 56.7% (227/400) and 23% (52/227), respectively. Cows (154/200) showed to be significantly more seropositive than calves (135/200). In qPCR, the number of calves and average quantification value (138/200; 1.3 × 10 6 ) A. marginale msp1α copies per μL proved to be higher when compared to that found for the cows (89/200; 3.9 × 10 4 ). The microsatellite analysis of the 26 sequences obtained from the msp1α gene revealed the presence of E (77%), C (15.4%) and B (7.7%) genotypes. Fourteen A. marginale strains were identified in the studied region, with eight that have never before been described in the literature (τ-10-13-13-18; τ-27-18; EV8-EV8-17; α-β-β-β-100; EV7-11-10-15; τ-11-11-27-18; τ-11-10-15; τ-27-13-18). Beef cattle are highly exposed to A. marginale in the Brazilian Pantanal. Moreover, a high genetic diversity of A. marginale, with eight new strains, was found in the studied region. While cows may act as chronic carriers, perpetuating the pathogen within the herd, male beef calves sold to other regions may disperse these strains., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

Anaplasma marginale is the most prevalent tick-borne pathogen in cattle in tropical and subtropical regions of the world. Major Surface Protein 1a (MSP1a) has been found to be a stable genetic marker for identifying A. marginale isolates within geographical regions. It is conserved in cattle during infection and tick-borne transmission of the pathogen. The aim of the present longitudinal study was to determine occurrences of genetic diversity associated with high prevalence of A. marginale under natural transmission conditions. Twenty calves were evaluated every 3 months during the first year of life. Rickettsemia levels due to A. marginale, measured as the number of msp1αcopies/ml in the blood of positive calves, ranged from 2.06×10(3) to 4.36×10(12). The numbers of MSP1a tandem repeats among MSP1a tandem repeats were 3 and 6. The predominant msp1α microsatellite was E, and another MSP1a tandem repeat was found that presented genotype G. Nineteen different MSP1a tandem repeats of A. marginale were found circulating in animals. The MSP1a tandem repeats 4-63-27 (27.5%), 78-24(2)-25-31 (n=21.6%) and τ-10(2)-15 (n=17.6%) were the ones most commonly observed. Twenty-two MSP1a tandem repeats resulted in new sequences with amino acid changes, as shown in this study. Thirty sequences were found for the first time in Brazil. Glycine, glutamate, serine and alanine amino acids were found at position 20. During the study, 80% (16/20) of the animals were infected by more than one genotype. Three animals were born infected, with MSP1a tandem repeats 4-63-27, 78-24(2)-25-31 and τ-10(2)-15, thus indicating occurrence of transplacental transmission. In the phylogenetic analysis, 19 different MSP1a tandem repeats of A. marginale were found in the cattle, which suggested that many MSP1a tandem repeats and high variation in MSP1a were occurring., (Copyright © 2015 Elsevier GmbH. All rights reserved.)