The Role of Transplacental Infection in Leptospira spp. Epidemiology in Cattle in Caatinga Biome, Brazil.

Leptospirosis is an infectious disease that affects domestic animals, wild animals, and humans. It represents a public health problem and has an important economic impact on livestock. This study aims to investigate the importance of genital and transplacental infection in the epidemiology of leptospirosis in cows maintained in Caatinga biome conditions, Northeastern Brazil, as well as reporting organs colonized by Leptospira spp. in embryos and fetuses. Blood, urinary tract (urine, bladder, and kidney), and reproductive tract (vaginal fluid, uterus, uterine tube, ovary, and placenta) samples were collected from 15 slaughtered pregnant cows. Two embryos and 13 fetuses were sampled. Central nervous system and choroid ovoid samples were collected from embryos. Blood, central nervous system, lung, peritoneal liquid, abomasal content, liver, spleen, urine, bladder, kidney, and reproductive system samples were collected from fetuses. Diagnostic methods included the microscopic agglutination test (MAT) using a collection of 24 serovars belonging to 17 different pathogenic serogroups of five species as antigens, as well as polymerase chain reaction (PCR). Anti-Leptospira spp. antibodies were found in 9 cows (60%), while 13 cows (86.67%) had at least one organ or urine with leptospiral DNA. No fetus was seroreactive. Among the embryos and fetuses, 13 (86.67%) presented leptospiral DNA, proving a high frequency of transplacental infection (100%). For cows, the most frequent biological materials regarding Leptospira spp. DNA detection were placenta (13 out of 15 samples; 86.7%), uterus (10 out of 15 samples; 66.7%), and vaginal fluid (5 out of 15 samples; 33.3%), while, for fetuses/embryos, the most frequent PCR-positive samples were choroid ovoid (1/2; 50%), spleen (6/13; 46.2%), kidney (5/13; 38.5%), and central nervous system (5/15; 33.3%). Sequenced samples based on the LipL32 gene presented 99% similarity with L. borgpetersenii. The results indicate that transplacental infection is an efficient way of spreading Leptospira spp. in cows maintained in Caatinga biome conditions. Therefore, prevention and control strategies must include actions that interrupt transmission through this alternative route.

Incorporating versus coating antimicrobials for polymethyl methacrylate: A systematic review.

STATEMENT OF PROBLEM: Incorporating and coating with antimicrobials are techniques that can confer antimicrobial action on polymethyl methacrylate (PMMA) denture bases, which can accumulate microorganisms and promote oral and systemic disease. PURPOSE: The purpose of this systematic review was to answer the question: "Do techniques for incorporating and coating antimicrobial agents in PMMA promote antimicrobial action?" MATERIAL AND METHODS: The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) checklist guidelines were followed, and the SCOPUS, PubMed/Medline, EMBASE, and Science Direct databases searched. The studies were selected in 2 stages, reading the titles and abstracts and then reading the selected studies in full. The risk of bias was analyzed by adapting the quasi-experimental studies tool by the Joanna Briggs Institute (JBI). RESULTS: A total of 970 articles were found in the databases; 71 were duplicates and, after reading the abstracts, 38 were selected for full reading. From these, 6 were excluded because they did not fulfill the inclusion criteria, and 32 studies were included in this review. Autopolymerizing, heat- polymerizing, and light-polymerizing resins were evaluated, with the incorporating technique prevailing over the coating, but both techniques effectively promoted antimicrobial activity. CONCLUSIONS: Incorporating and coating antimicrobial agents are effective methods of promoting antimicrobial activity in PMMA. Combining the 2 methods led to increased antimicrobial activity compared with each individually.

Strong Evidence of the Role of Donkeys in the Epidemiology of Leptospira spp. in Semiarid Conditions.

Donkeys (Equus asinus) are historically known for their close relationship to humanity, which raises the need to study zoonotic diseases that affect them. In this perspective, leptospirosis stands out as a disease with an economic and public health impact, and its occurrence is facilitated in times of higher rainfall indexes, especially in large urban centers. In view of the scarcity of information about leptospirosis in donkeys, the objective of this study was to detect the presence of Leptospira spp. and anti-leptospiral antibodies in donkeys rescued by a zoonosis center located in the Caatiga biome, Brazilian semiarid region. Overall, 30 donkeys of both sexes, aged between 4 months and 15 years, were used, from which 64 serum samples were collected and submitted to the microscopic agglutination test (MAT). In addition, 64 samples of urine, vaginal and preputial fluid, in duplicates, were subjected to the polymerase chain reaction (PCR) and microbiological. Sixteen (53.3%) animals tested positive in at least one diagnostic test, 12 (40%) of which were positive at MAT and seven (23.3%) in the molecular and bacteriological detection (urine, vaginal, and preputial fluid samples). This is the first report identifying donkeys infected with Leptospira spp. by molecular and bacteriological diagnosis in Brazil, and the first in the world to detect this agent in their genital fluids. The study also shows that donkeys are commonly exposed to leptospires in the Caatinga biome, and this constitutes a One Health-based concern, demonstrating the importance of broad studies where large numbers of humans and animals coexist when investigating zoonotic infections and when planning and implementing control measures for donkeys-associated leptospirosis.

Antiplasmodial activity of coumarins isolated from Polygala boliviensis: in vitro and in silico studies.

Polygala boliviensis is found in the Brazilian semiarid region. This specie is little chemically and biologically studied. Polygala spp. have different metabolites, especially coumarins. Studies indicate that coumarins have antimalarial potential, denoting the importance of researching new active compounds from plants, since the resistance of Plasmodium strains to conventional therapy has increased. The present study aimed to evaluate the antiplasmodial activity of auraptene and poligalen against a chloroquine-resistant strain of Plasmodium falciparum. Coumarins were isolated from P. boliviensis by open column chromatography and identified by Nuclear Magnetic Resonance Spectroscopy. A cytotoxicity assay was carried out using MTT test, and the in vitro antiplasmodial activity was evaluated using the W2 strain. The antiplasmodial activity results found were IC50=0.171 ± 0.016 for auraptene and 0.164 ± 0.012 for poligalen; the selectivity indexes were 78.71 and 609.76, respectively. Inverse virtual screening in the BRAMMT database by OCTOPUS 1.2 was applied to coumarins to find potential P. falciparum targets and showed higher affinity energy of auraptene for purine nucleoside phosphorylase (PfPNP) and of poligalen for dihydroorotate dehydrogenase (PfDHODH). Molecular Dynamics studies (MD and MM-GBSA) approach were applied to calculate binding energies against selected P. falciparum targets and showed that all coumarins were stable at the binding site during simulations. Furthermore, energies were favorable for complexation. This is the first report of auraptene in P. boliviensis species and of in vitro antiplasmodial activity of auraptene and poligalen. In silico studies indicated that the mechanism of action of coumarins is the inhibition of PfPNP and PfDHODH.Communicated by Ramaswamy H. Sarma.

Detection of Leptospira spp. in genitourinary tract of female goats managed in the brazilian semiarid.

Studies related to the prevalence of leptospirosis in the semiarid region showed that even during long periods of drought, the disease has a remarkable frequency in herds in the region. It is a neglected disease and the extent of its effects in the Brazilian semiarid region is not known. The dynamics of this agent is well studied in the urinary tract, however, there are not many studies regarding the genital tract in female goats. Observing this scenario, the present work aimed to diagnose Leptospira spp. in female goats kept in the Brazilian semiarid region by means of serological, molecular and isolation techniques. Blood samples, vaginal fluid, urine and fragments of organs from the genitourinary tract were collected from 40 goats destined for slaughter. Microscopic agglutination test (MAT) was used as a serological technique, with a battery of 24 serovars. The Polymerase Chain Reaction (PCR) of the vaginal fluid, urine and organ fragments was performed, as well as the bacterial growth of these same products in a selective medium. Isolation positive samples were subjected to PCR. It was observed that two (5%) animals were serologically positive for the Pyrogenes serogroup. A total of 29 (72.5%) animals were PCR positive, with DNA present in 51/160 (31.8%) samples from the genital tract and 34/120 (28.3%) from the urinary tract, with no statistical difference. For bacterial growth, 22/40 (55%) animals were positive for growth, with morphology being observed in 19/160 (11.8%) for the genital tract and 16/120 (13.3%) for the urinary tract, with no statistical difference. Two uterus samples showed 99% similarity with L. interrogans after sequencing. Thus, female goats kept under semiarid conditions were positive for Leptospira spp, with positive samples from both the urinary and genital tracts, which possible is an alternative way of adapting and maintaining the agent for severe and adverse conditions.

Chemical composition of selected insect meals and their effect on apparent total tract digestibility, fecal metabolites, and microbiota of adult cats fed insect-based retorted diets.

Insect meals are novel and potentially sustainable protein sources. The objectives of this study were to determine the chemical composition and standardized amino acid digestibility using the cecectomized rooster model of three selected insect meals (i.e., speckled cockroach [SC], Madagascar hissing cockroach [MC], and superworm [SW]) and to determine the effects of these insect meals on food intake, apparent total tract digestibility (ATTD) of macronutrients, fecal scores, and metabolites of adult cats fed insect- or chicken-based retorted diets. This study consisted of a complete randomized design, with 28 adult cats randomly assigned to one of the four experimental retorted diets: Control (chicken-based diet), SC diet, MC diet, or SW diet. All animal procedures were approved by the University of Illinois Institutional Animal Care and Use Committee. All diets were formulated to be complete and balanced and meet or exceed the nutritional requirements of adult cats. The experimental period was 28 d, with the first 7 d allotted for diet adaptation. The total fecal collection was completed during the last 4 d of the experimental period. On day 21, a fresh fecal sample from each cat was collected for the determination of fecal metabolites and microbiota. Food was offered twice daily to maintain body weight and body condition score. Among the three selected insect meals evaluated, oleic acid, palmitic acid, linoleic acid, and stearic acid were the most prevalent fatty acids. Branched-chain amino acids and arginine were the most preponderant indispensable amino acids in these insect meals. ATTD of dry matter, organic matter, acid-hydrolyzed fat, and crude protein did not differ among treatments (P > 0.05), and all diets were well digested by the cats. Similarly, fecal scores did not differ among the treatments and were within ideal range. No differences (P > 0.05) in fecal metabolite concentrations or microbiota diversity were observed among cats fed different experimental diets; only a few genera from Firmicutes and Bacteroidota phyla differ (P < 0.05) in cats fed SW diet in contrast to other dietary treatments. In conclusion, the selected insect meals evaluated herein are rich in linoleic acid, an essential fatty acid for cats. Insect-based retorted diets led to comparable results to those achieved with a chicken-based retorted diet, suggesting that these novel protein sources might be adequate alternative ingredients in feline diets.

Insect meals are novel and potentially sustainable protein sources. The objectives of this study were to determine the nutrient composition of speckled cockroach, Madagascar hissing cockroach, and superworm (SW) and to determine the effects of these insect meals on food intake, digestibility of macronutrients, fecal scores, metabolites, and microbiota of adult cats fed insect- or chicken-based wet pet foods. Among the three selected insect meals evaluated, oleic acid, palmitic acid, linoleic acid, and stearic acid were the most prevalent fatty acids. Branched-chain amino acids and arginine were the most preponderant indispensable amino acids in these insect meals. All diets were well digested by the cats with no differences observed on macronutrient digestibility. Similarly, fecal scores did not differ among the treatments and were within the ideal range. No differences in fecal metabolite concentrations were observed. Only a few genera from Firmicutes and Bacteroidota phyla differ in cats fed SW diet in contrast to other dietary treatments. Overall, the selected insect meals evaluated herein are rich in linoleic acid, an essential fatty acid for cats. Insect-based retorted diets led to comparable results to those achieved with a chicken-based retorted diet, suggesting that these novel protein sources might be adequate alternative ingredients in feline diets.

Cross-sectional survey for sheep leptospirosis in the northeast region of Brazil.

Sheep farming has shown expressive growth, even in the Northeast region of Brazil, with rainfall below expectations in the last decade. This growth has been observed not only in the number of herds, but also in the number of properties involved in sheep farming. Although expressive, the production of small ruminants still presents low levels of performance, mainly due to the occurrence of diseases, which constitute an obstacle to this activity. The aim of this study was to determine the factors associated with the seroprevalence of leptospirosis in sheep in Northeastern Brazil. Blood samples were collected from 3438 sheep in 190 herds from 48 municipalities, belonging to the states of Ceará, Paraíba, Piauí, Rio Grande do Norte and Sergipe. Serological diagnosis was performed using the microscopic agglutination test (MAT). In 76.3 % (95 % CI = 69.79-81.81) of the herds there was at least one animal seropositive for at least one of the Leptospira spp. serogroups and 14.94 % (95 %CI = 13.8-16.18) of the animals were seropositive. The state of Paraíba had the highest herd (95.7 %) and animal (26.6 %) prevalences, followed by Ceará (93.9 % and 23.7 %, respectively). The most frequent serogroups were Autumnalis (18.68 %), Australis (17.9 %) and Pyrogenes (15.18 %). There was no association between the studied variables and the herd-level prevalence of leptospirosis. High herd and animal-level seroprevalences was found for sheep leptospirosis in the semiarid region of Northeastern Brazil, and there is a need to implement control and prevention measures, such as quarantine, serology, vaccination and adequate treatment, in addition to the introduction of proven healthy animals, mainly in the states of Paraíba and Ceará, aiming to reduce the prevalence of disease.

Oral Angiotensin-(1-7) Peptide Modulates Intestinal Microbiota Improving Metabolic Profile in Obese Mice.

BACKGROUND: Obesity is a serious health problem that dysregulate Renin-Angiotensin System (RAS) and intestinal microbiota. OBJECTIVE: The present study aimed to evaluate the Angiotensin-(1-7) [ANG-(1-7)] oral formulation effects on obese mice intestinal microbiota. METHODS: Mice were divided into four groups: obese and non-obese treated with ANG-(1-7) and obese and non-obese without ANG-(1-7) during four weeks. RESULTS: We observed a significant decrease in the fasting plasma glucose, total cholesterol, triglycerides, and Low-density lipoprotein levels and increased High-density lipoprotein in animals treated with ANG-(1-7). The histological analysis showed intestinal villi height reduction in mice treated with ANG-(1-7). Additionally, increased Bacteroidetes and decreased Firmicutes (increased Bacteroidetes/ Firmicutes ratio) and Enterobacter cloacae populations were observed in the High-Fat Diet + ANG-(1-7) group. Receptor toll-like 4 (TLR4) intestinal mRNA expression was reduced in the HFD+ANG-(1-7) group. Finally, the intestinal expression of the neutral amino acid transporter (B0AT1) was increased in animals treated with ANG-(1-7), indicating a possible mechanism associated with tryptophan uptake. CONCLUSION: The results of the present study suggest for the first time an interaction between oral ANG-(1-7) and intestinal microbiota modulation.

Diet Supplementation with Madagascar Cockroach Flour (Gromphadorhina portentosa) Improved Malnourished Mice Metabolism and Ameliorated Liver Inflammatory Markers.

BACKGROUND: Malnutrition and accessible high-quality protein food sources are two of the world's alimentary challenges. Edible insects are nowadays recognized as a possible functional food solution with lower environmental impacts and beneficial health effects. OBJECTIVE: In this context, the aim of the present study is to evaluate Madagascar cockroach (Gromphadorhina portentosa) flour supplementation effects on a malnourished mice model, considering its effects on metabolism, adiposity, and inflammatory liver profiles. METHOD: Male Swiss mice are divided into five groups and fed with experimental diets for eight weeks, including a standard diet (ST) ad libitum, AIN93 ad libitum (AIN), insect flour-enriched AIN93 (AIN+IM), AIN-40% feed restriction (AIN-FR), and insect flour-enriched AIN-40% of feed restriction (AIN-FR+IM). The metabolic profiles, adipose tissue, biochemical parameters, and liver IL-6 and IL-10 expression are evaluated. RESULTS: The main findings show a body weight and metabolism improvement followed by an increased recovery of the adipocyte area in the AIN-FR+IM group when compared to the AIN-FR malnourished group. Reduced hepatic IL-6 and increased IL-10 expression are also detected in the AIN-FR+IM group. CONCLUSION: The results show that insect flour supplementation enhances both body weight and adiposity gain/recovery. The results also show hepatic improvement of inflammatory markers.

Serological, molecular and bacteriological approaches for detecting Leptospira sp. carrier rams maintained in semiarid conditions.

Even in the adverse environmental conditions of the semiarid region, leptospires can survive and spread by alternative routes of transmission, such as sexual in ewes, however, there is no data on rams. Thus, the present study aimed to evaluate the use of serological, molecular and microbial tools applied to diagnosis of Leptospira sp. Infection in rams maintained in semiarid conditions. Biological samples of urinary (urine, kidney and bladder) and genital (vas deferens, epididymis tail and vesicular gland) tracts were collected from 40 slaughtered rams for polymerase chain reaction (PCR) and bacterial isolation, as well as blood samples for antibody detection through microscopic serum agglutination test (MAT). Anti-Leptospira antibodies were found in five (12.5%) animals with antibody titer of 50 and 2 (5%) for the titer 100 for serogroups Pyrogenes, Ballum, Icterohaemorrhagiae and Australis. Leptospira sp. DNA was found in PCR of organs and urine of 30 (75%) animals. Overall, 240 fragments of organs from the urinary and genital tracts and urine were evaluated, with 93 (38.7%) positive samples, being 48/120 (40%) for the urinary tract and 45/120 (37.5%) for the genital. There was no statistically significant difference between the tracts. A bladder sample was sent for sequencing and showed 99% similarity with L. interrogans. Of the 240 cultures evaluated, 59 (24.5%) had leptospire growth, being that 23 (39%) were confirmed in PCR. Considering the PCR of organs and urine and bacterial growth as gold standards, the cut-off 50 in MAT showed greater sensitivity when compared to cut-off 100, regardless of the material used. The great proportion of leptospiral DNA in organs, urine and culture and bacterial growth from the genital tracts reinforce its importance as an extra-renal site and highlights the possible role of rams in venereal transmission, as well as the sensitivity of the cut-off 50 suggested its adoption in the serology of rams maintained in semiarid conditions.

Visualization and Manipulation of Bilayer Graphene Quantum Dots with Broken Rotational Symmetry and Nontrivial Topology.

Electrostatically defined quantum dots (QDs) in Bernal stacked bilayer graphene (BLG) are a promising quantum information platform because of their long spin decoherence times, high sample quality, and tunability. Importantly, the shape of QD states determines the electron energy spectrum, the interactions between electrons, and the coupling of electrons to their environment, all of which are relevant for quantum information processing. Despite its importance, the shape of BLG QD states remains experimentally unexamined. Here we report direct visualization of BLG QD states by using a scanning tunneling microscope. Strikingly, we find these states exhibit a robust broken rotational symmetry. By using a numerical tight-binding model, we determine that the observed broken rotational symmetry can be attributed to low energy anisotropic bands. We then compare confined holes and electrons and demonstrate the influence of BLG's nontrivial band topology. Our study distinguishes BLG QDs from prior QD platforms with trivial band topology.

Coelioscopic-assisted prefemoral ovariosalpingectomy in a d`Orbigny slider (Trachemys dorbigni) using a digital otoscope.

The purpose of this study was to assess the unilateral prefemoral coelioscopic-assisted approach for ovariosalpingectomy in a d`Orbigny slider (Trachemys dorbigni) using a digital otoscope. Twenty healthy turtles were randomly assigned to one of two groups, for right (GR; n=10) or left (GL; n=10) prefemoral access, for coelioscopic-assisted ovariosalpingectomy. Anesthesia and surgery times, body weight, and ovary/oviduct weight data were recorded. Anesthesia and surgery times did not differ significantly between the groups. Wound closure was the most time-consuming surgical step. Ovary and body weights significantly affected the exposure time of the ipsilateral and contralateral ovaries, respectively. Two intraoperative complications were reported. All the animals recovered uneventfully. The digital otoscope can be safely and effectively used for coelioscopic-assisted single-access, unilateral prefemoral ovariosalpingectomy in d`Orbigny slider.

Didelphis albiventris as a carrier of Leptospira sp. in the central nervous tissue in the semiarid region of Northeast, Brazil.

Leptospirosis has been investigated in several species of wild animals. The white-eared opossum (Didelphis albiventris) is a mammal common in the brazilian semi-arid, so, this study aimed to investigate its role in the occurrence of the leptospirosis in the region Northeast of Brazil. 12 animals were used, from which samples were collected for the attempt of isolation, molecular detection and serological examination. There was no microbial growth, nor were any anti-Leptospira sp. antibodies found in the serological samples. The PCR detected leptospiric DNA in the central nervous system (CNS) of five animals (41.7 %). The gene in one of the samples was sequenced and showed identity with Leptospira interrogans. The presence of Leptospira sp. in the CNS of Didelphis albiventris does not allow the characterization of the studied animals as reservoirs with potential for transmission of the pathogen in the region, however it represents a site that needs to be further investigated.

Leptospira sp. vertical transmission in ewes maintained in semiarid conditions.

Leptospirosis is a globally distributed disease associated with reproductive failures in livestock; however, its pathogenesis has not been fully elucidated. Results from the present study indicate there is a presence of Leptospira sp. in organs and fluids of fetuses from ewes slaughtered in the semiarid region of Brazil. Twenty-nine fetuses from 23 ewes determined to be Leptospira sp.-positive using PCR were sampled (14 and 15 in dry and rainy seasons, respectively). Fetal samples of blood, central nervous system (CNS), lung, liver, spleen, stomach contents, peritoneal fluid, kidney, bladder, urine and reproductive system were collected. Diagnostic methods included the microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Of the 29 fetuses, 24 (82.8 %) had at least one Leptospira sp.-positive organ or fluid, as determined using PCR, and of a total of 209 samples, 62 (29.7 %) contained leptospiral DNA. Of the 99 samples collected during the dry season, 42 (42.4 %) were positive, and of 110 samples collected during the rainy season, 20 (18.2 %) were positive (P = 0.0001). There was deoxyribonucleic acid (DNA) sequencing of three samples of kidney, CNS and liver, and in all of these, there was 99.3 % similarity with Leptospira interrogans. Leptospires were present in cultures of pooled samples from fetuses with deformities. Results indicate there is vertical (maternal-to-fetus) transmission which would represent an alternative transmission route for the spread of Leptospira sp. in ewes, suggesting molecular detection is essential in the investigation of leptospirosis in fetuses to identify animals that have been infected with this bacterium.

Use of serological and molecular techniques for detection of Leptospira sp. carrier sheep under semiarid conditions and the importance of genital transmission route.

It is possible that there are peculiarities in the epidemiology of leptospirosis in regions with a semiarid climate, where the environment is often adverse, allowing the occurrence of alternative transmission routes. The objective of the work was to generate contributions to the diagnosis and epidemiology of Leptospira sp. infection in sheep reared in semiarid conditions, using serological, molecular and microbiological techniques for diagnosis in dry and rainy seasons. Samples of blood, vaginal fluid, urine, bladder, kidney, uterus, uterine tube, ovary and placenta were collected from 104 sheep (52 animals per season - dry and rainy) slaughtered in the Brazilian semiarid. Diagnostic tests performed were microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Anti-Leptospira sp. antibodies were found in 26 (25%) of the animals analyzed by MAT at 1:50 dilution (cut-off 50), while 69 (66.3%) animals had at least one organ/fluid with the presence of Leptospira sp. DNA. Overall, PCR was performed on 758 fragments of organs/fluids from the genital and urinary tracts, and 519 (68.5%) samples tested positive. PCR-positivity was statistically different in dry (46.2%) and rainy (11.5%) seasons for vaginal fluid. It was possible to perform the DNA sequencing in nine samples with 99% similarity to L. interrogans and recovery of viable strains in three samples of vaginal fluid. Regardless of the biological material used in PCR to detect carrier animals and the season, the highest MAT sensitivity values were obtained with cut-off 50 compared to 100. The results obtained indicate that, even in the adverse environmental conditions of the semiarid region, leptospires may survive and propagate by alternative routes of transmission, such as sexual, and the presence of PCR-positive genital tracts in ewes suggests that sexual transmission may play an important role in the epidemiology of the disease in sheep in Brazilian semiarid. In addition, it is suggested the use of titer 50 as cut-off point at serology in semiarid conditions.

High frequency of seropositive and carriers of Leptospira spp. in pigs in the semiarid region of northeastern Brazil.

Carrier animals are considered key in the transmission cycle of leptospirosis. Although investigations have been carried out on several species, the role of pigs in the epidemiology of the disease is still poorly studied in the semi-arid region. Thus, the objective of this study was to determine the presence of Leptospira spp. in the genitourinary tract of pigs intended for slaughter. Fifty pigs were used: adults and unvaccinated. Samples of the kidney, urine, and vaginal fluid were collected for the molecular detection of Leptospira spp. and blood samples for the serological test. The molecular test was performed using the polymerase chain reaction (PCR), and the serological test was performed with the microscopic agglutination test (MAT). Samples with DNA amplification were submitted to genetic sequencing. Twenty (40%) animals were found with anti-Leptospira spp. antibodies, and the majority of the reactions (50%) occurred for the serogroup Tarassovi. Leptospiral DNA was found in the tissue of 11 (22%) pigs. The gene from a urine sample was sequenced and showed similarity to L. borgpetersenii. The results evidenced a high rate of porcine carriers; therefore, they appear to be important sources of agent infection, being potential transmitters of the disease to other animal species and man.

Influence of breed on the clinical and hemato-biochemical parameters in sheep experimentally infected with Leptospira sp.

Early diagnosis of leptospirosis may aid in a favorable prognosis in infected animals, but there are few reports of clinical and hematochemical changes in the ovine species, nor whether the breed exerts any influence on the response to infection. Thus, the objective of the present study was to evaluate the clinical, biochemical and hematological alterations in Santa Inês and crossbred ewes challenged with Leptospira interrogans serogroup Pomona serovar Kennewicki. Twenty-four sheep were used in this experiment, 12 crossbred (group A) and 12 Santa Inês (group B). In each group, sheep were conjunctivally and intraperitoneally challenged. During 60 days post-infection the ewes were evaluated for the presentation of clinical signs and the blood was collected for hemogram and serum biochemistry. Concentration of urea and creatinine; serum aspartate aminotransferase activity (AST); gamma-glutamyltransferase (GGT); total protein and albumin; total bilirubin, direct and indirect were analyzed. The urine of these sheep was collected for urinalysis. Only two Santa Inês sheep showed blood in the urine. Clinical signs implicated in Leptospira sp. infection were not identified. Some sheep had anemia, especially crossbred. However, anemia may be attributed to the more effective cellular response that has been identified in crossbred animals. Only one animal presented leukocytosis with neutrophilia, while 11 presented atypical leukopenia, especially those of the Santa Inês breed. There was a decrease in total protein and albumin, as well as the increase in gammaglutamyltranspeptidase (GGT), especially in Santa Inês sheep. The level of aspartate aminotransferase (AST) remained within the normal range for the species. A significant increase (p < 0.05) in conjugated bilirubin levels in challenged animals was detected. Only one sheep intraperitoneally challenged presented a high level of urea in the blood, but the creatinine level remained within the normal range. The intraperitoneal route was responsible for more significant changes (p < 0.05) in the hemogram and biochemistry when compared to the conjunctival route. The results indicate that crossbred sheep have a more efficient cellular response than Santa Inês sheep, which may confer a greater resistance to infection. Clinical signs are not good parameters to follow the development of leptospirosis in crossbred and Santa Inês breed. Hematological and biochemical analyzes were useful in the detection of anemia and possible liver changes caused by leptospirosis. The intraperitoneal route was able to cause more conclusive alterations of the analyzed parameters, however, it is possible that the alterations caused by the conjunctival route reproduce in a more faithful way what happens in a natural situation of infection.

Microbiological, molecular, and histopathological findings in goats experimentally infected with Actinobacillus seminis.

The objective of this study was to experimentally evaluate the pathogenicity of an Actinobacillus seminis isolate named SAAS01 in goats. Animals were challenged with 2 mL of a suspension containing 1,5 × 108 CFU/mL of A. seminis (SAAS01 isolate) through the intrapreputial, epididymis tail, and conjunctival routes. Epididymis and testicular fragments were submitted to histopathological exam, and semen samples underwent microbiological and molecular diagnoses. Clinically, a unilateral increase in firm consistency was observed in the epididymis and testicles of two animals inoculated in epididymis tail and in one animal inoculated through conjunctival sac; this firmness continued until the day of euthanasia. Two goats inoculated through epididymis tail and conjunctival sac routes presented histopathological findings with macroscopically and microscopically significant changes. A. seminis was isolated from semen samples collected from goats inoculated through the epididymis tail and conjunctival sac routes. A. seminis DNA was amplified from six semen samples of three goats inoculated through the epididymis tail, two in conjunctival sac and one through intrapreputial route. The experimental infection model using goats confirmed the pathogenicity of the A. seminis isolate, demonstrating the predilection of the agent for the epididymis, with clinical signs, histopathological lesions, bacterial isolation, and a positive molecular diagnosis.

Strategies of the control of an outbreak of leptospiral infection in dairy cattle in Northeastern Brazil.

The aim of the present study was to describe the strategies of the control of an outbreak of leptospiral infection in dairy cattle in Maranhão State, Northeastern Brazil. In the period from January to July 2015, 18 (17%) out of 106 cows presented abortion, six (5.7%) stillbirth, and 12 (11.3%) repeated estrus, totaling 24 animals with reproductive problems. The diagnosis of leptospirosis was based on serology (microscopic agglutination test-MAT), bacteriological culture, and polymerase chain reaction (PCR). Antibiotic therapy, vaccination protocols, and changes in management practices were suggested as control measures. Of all animals on the farm (n = 280), 136 (48.6%) were seropositive for at least one serovar of Leptospira sp. No pure leptospiral culture was obtained. Eight of the animals with reproductive problems yielded positive PCR results (vaginal fluid of seven animals and urine and vaginal fluid of one animal). Genetic sequencing of a vaginal fluid/urine PCR-positive sample revealed Leptospira borgpetersenii. One year after the adoption of control measures, no reproductive problems were observed. Thus, leptospirosis probably caused the reproductive failures in the herd, and the control and prevention measures implemented were efficient in controlling the disease.

Susceptibility among breeds of sheep experimentally infected with Leptospira interrogans Pomona serogroup.

Leptospirosis is a disease that negatively affects the productive and reproductive indices of ruminants. Sheep are considered highly resistant to infection, although susceptibility may vary among breeds. Thus, the aim of the present study was to analyze the susceptibility between sheep breeds to the experimental infection by leptospires of the Pomona serogroup. Pomona serogroup, Kennewicki serovar strain (1 × 107 bacteria) was inoculated via the conjunctival route in 12 sheep divided into two groups, one comprising Santa Inês ewes and the other comprising crossbred sheep. In each group, five ewes were challenged with the bacterial strain and one was used as control. All sheep were monitored for 60 days, during which blood samples were collected for serological diagnosis and urine and vaginal fluid samples for molecular and microbiological analyses. Finally, as ewes were submitted to euthanasia and necropsy, some tissues of interest were collected for microbiological, molecular, and histopathological diagnoses. The groups were compared regarding the number of positive reactions according to diagnostic tests. All sheep in each group presented antibodies to Leptospira in all serological analyses, except animals of the control group. However the Santa Inês sheep presented higher concentration and duration of the titers, and their positive reactions were detected earlier than those in crossbred sheep. The antibody titers in group A (median 200, geometric mean 317.48) were significantly different from the group B (median 800, geometric mean 918.96) at D60 post-infection (P = 0,032). The Santa Inês sheep presented a higher number of positive reactions than did the crossbred sheep in the molecular diagnostic tests. According to the molecular diagnosis, the Santa Inês sheep presented more reactions (urine and vaginal fluid) compared to crossbred ewes, but there was no predominance in the detection of leptospiral DNA when comparing urine and vaginal fluid results, nor even between the number of positive kidneys and uterus. The Santa Inês sheep presented a higher number of positive bacteriological cultures. No sheep in either group presented alterations in anatomopathological and histopathological findings. Pure-bred sheep may be more susceptible than crossbred sheep to infection by Leptospira sp. Our findings emphasize the importance of the genital tract as a site of extraurinary infection and indicate the possibility of venereal transmission in the species.