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Purpureocillium lilacinum is a filamentous and hyaline fungus cosmopolitan, saprophytic, largely used in the biological control of plant-parasitic nematodes and insects, also considered an emerging and opportunistic human pathogen. The standard treatment for hyalohyphomycosis caused by P. lilacinum is not yet defined, since this fungus is resistant to different antifungals, in vitro and in vivo. The aim of this study was to evaluate and compare in vitro antifungal activity against environmental and clinical P. lilacinum isolates and our results demonstrated that these isolates can be resistant to newer generation triazoles, such as voriconazole, and to caspofungin, a drug of the echinocandin class. In summary, we highlight the importance of knowing the different susceptibility profiles of P. lilacinum isolates, and besides that, the emergence of uncommon human and animal opportunistic fungi, such P. lilacinum, especially during COVID-19, highlight the need for antifungal susceptibility testing of isolates since empirical therapy with different treatment schedules failed in great number of patients., (© 2022 The Society for Applied Microbiology.)

Objective: Our investigation was conducted to evaluate the incidence of umbilical pathologies and the result of related surgical interventions in Sicilian cattle., Study Design: 320 (214 females, 106 males) cattle with umbilical lesions were collected, anesthetized, operated on and follow-up recorded. We evaluated the injury incidence rate., Population: The animals considered were: high productivity dairy cattle (Holstein Friesian and Brown Swiss); meat breeds (Charolaise and Limousine) and some crosses., Results: The highest injury rate was simple omphalocele, followed by purulent omphalitis. A high percentage was found in young cattle. Friesians are the most prone to navel diseases. The dairy breeds, compared to the meat breeds, and double aptitude and crosses, were the most affected. The most common was simple omphalocele, with a small hernial port (1 to 3 cm), while the rest was greater than 3 cm, with severe organs or complications., Methods: the most commonly used surgery was herniorraphy with autologous reinforcement, while classic sutures and mesh sutures were used in the remaining cases. Follow-up demonstrated healing in most of the treated subjects., Conclusions: The study aimed to estimate some surgical clinical cases in Sicilian cattle. Most injuries are lower abdomen injuries, mostly simple or complicated umbilical hernias. Young dairy females were the most affected., Meaning/impact: Radical surgery is the treatment of choice in the vast majority of symptomatic or asymptomatic umbilical diseases; moreover, the correct management and the choice of an appropriate surgical approach allow to obtain an effective treatment of the lesions., Simple Summary: In cattle farm, the correct management of births is a fundamental step for the future of the breeding. In fact, there are numerous critical factors in this phase: calving area management, umbilical disinfection, correct administration of colostrum, management of individual pens. Generally, after a few hours from birth, the calf is transferred in individual pens, the walls of this box allow visual and tactile contact but avoid the possibility of mutual sucking of the umbilical region, a risk factor, in that area, of infections and hernias. In the study have been collected umbilical interventions carried out over twenty years by a team operating in south-eastern Sicily: Ragusa. Out of a total of 320 cases, have been reported: the prevalence of types of umbilical tumefactions, anesthetic techniques and surgical techniques performed. The purpose of the work was to highlight any predispositions of race, sex, age regarding the lesion considered and above all, the effectiveness of the surgical therapy., Competing Interests: The authors declare no conflict of interest., (© 2022 The Authors. Published by Elsevier Ltd.)

Curraleiro Pé-Duro (CPD) cattle, a Brazilian local breed, are animals that are highly resistant to infectious and parasitic diseases. Strategies for the conservation of the breed and the genetic resistance to diseases should also consider the characteristics of the breed. The objective of this study was to analyze the diversity and population structure of the CPD breed using microsatellite markers and to correlate the serological profiles for causative agents of brucellosis, leptospirosis, neosporosis, leukosis, infectious bovine rhinotracheitis, and bovine viral diarrhea. DNA samples of 608 bovines were amplified and genotyped using 28 microsatellite markers for breed characterization. The genotypes were assigned to three clusters, indicating a substructure population related to the geographic distance. The observed heterozygosity was lower than that expected in most loci, and fixation index (Fst) in the subpopulation values ranged from 0.03073 (ETH3) to 0.402 (INRA63) on cluster 1, from 0.00 (INRA32) to 0.39359 (INRA63) on cluster 2, and from 0.00 (INRA32) to 0.28483 (TGLA53) on cluster 3. The Brucella spp ., Neospora caninum , and Bovine Leukemia Virus frequencies were significantly different ( p < 0.05) between clusters. The differences between the occurrences of diseases were not sufficient to indicate a genetic subpopulation with increased resistance to infections., 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 © 2022 Freitas, Dias, Veríssimo, Lobo, Costa, Moura, Landi, Martínez, Carmo and Fioravanti.)

Malaria is an acute febrile disease caused by a protozoan of the genus Plasmodium . Light microscopy (LM) is the gold standard for the diagnosis of malaria. Despite this method being rapid and inexpensive, it has a low limit of detection, which hampers the identification of low parasitemia infections. By using multicopy targets and highly sensitive molecular techniques, it is possible to change this scenario. In this study, we evaluated the performance of droplet digital PCR (ddPCR) to detect Plasmodium DNA obtained from saliva samples (whole saliva and buccal swab) of 157 individuals exposed to malaria transmission from the Brazilian Amazon region. We used the highly sensitive ddPCR method with non-ribosomal multicopy targets for Plasmodium vivax (Pvr47) and Plasmodium falciparum (Pfr364). There was good concordance between the quantitative real-time PCR (qPCR) results from the saliva and blood, except for mixed-species infections. The sensitivity of qPCR was 93% for blood, 77% for saliva, and 47% for swabs. Parasite DNA was not detected in saliva samples in low-density infections compared with the detection in blood samples. ddPCR showed increased sensitivity for detecting Plasmodium in the blood and swabs (99% in blood, 73% in saliva, and 59% in swabs). Notably, ddPCR detected more mixed infections in the blood (15%), saliva (9%), and swabs (18%) than qPCR. Our data showed that the differences between ddPCR and qPCR were the result of a higher number of P. falciparum infections detected by ddPCR. Overall, there was a moderate correlation between parasite densities estimated by the different methods in the blood. Our findings highlight the possibility of using non-invasive sample collection methods for malaria diagnosis by targeting multicopy sequences combined with highly sensitive molecular methods., Competing Interests: The reviewer JL-J declared a shared affiliation with the authors GC, DA, LC, CF, TN at the time of the review. The remaining 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 © 2022 Costa, Alvarenga, Aguiar, Louzada, Pereira, de Oliveira, Fonseca Júnior, Carvalho, Ferreira Alves de Brito and Nóbrega de Sousa.)

This study was carried out to investigate the frequency of genetic variants related to body mass index (BMI) and type 2 diabetes (T2D) and evaluating the potential impact of risk alleles on susceptibility to these disorders in six indigenous peoples from Brazilian Amazon region. The majority of Fst values for pairwise population comparisons among the indigenous groups are low or moderate. The indigenous people show high values of differentiation with Africans, Europeans and Southeast Asians and moderate values with East Asian and American populations, as expected. The allelic frequencies among indigenous indicate that the majority of associations observed with T2D in continental populations can be replicated in native Amazonians. The genetic risk scores calculated for T2D in indigenous are high and similar to those calculated for Americans and East Asians, while the estimates obtained for obesity are low, probably due to the low frequencies of the risk allele of the FTO gene found in our samples. ADRB3-rs4994 and ABCC8-rs1799854 genes showed a significant association with BMI and waist circumference, and the KCNJ11-rs5219 gene with hyperglycemia. These results emphasize the importance of knowing the genetic variability underlying complex genetic diseases in indigenous peoples and the search for particular or rare variants.

Human T-lymphotropic viruses 1 and 2 (HTLV-1 and HTLV-2) are retroviruses that originated on the African continent and dispersed throughout other continents through human migratory flows. This study describes the prevalence of HTLV-1 and HTLV-2 infection in residents of 11 quilombo remnant communities in the state of Pará, Brazil, and the associated risk factors. A total of 859 individuals (334 men and 525 women), aged between 7 and 91 years, participated in the study. All subjects answered a questionnaire with questions on sociodemographic characteristics and on risk factors associated with HTLV infection, and blood samples were collected and separated into plasma and leukocytes. An immunoenzymatic assay (ELISA; Murex HTLV-I+II, DiaSorin, Dartford, UK) was used as a screening test, and positive samples were subjected to line immunoassay confirmatory tests (Inno-LIA HTLV I/II Score FUJIREBIO) and DNA extraction for subsequent real-time PCR to differentiate the viral type. Four of the 859 individuals were seropositive for HTLV. HTLV-1 infection was confirmed in one individual from the Itamoari community (0.92%), and HTLV-2 infection was confirmed in two individuals from São Benedito (3.17%) and in one individual from Arimandeua (2.22%). Blood transfusion was the only risk factor associated with HTLV infection in this study. This study reports the occurrence of HTLV-1 and HTLV-2 in quilombo remnant communities in the state of Pará. Considering the African origin of the virus and its introduction into Brazil from the slave trade, the continued evaluation of quilombola communities in the state of Pará is essential to better characterize the distribution of infections in these populations and to create public health policies for the control of the spread of the virus and associated diseases., 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 © 2022 Brito, Cardoso-Costa, Roland Junior, Pereira, Lopes, Santos, de Lima, Abreu, Lima, Lima, Cayres Vallinoto, Santos, Guerreiro and Vallinoto.)

Early diagnosis and treatment are fundamental to the control and elimination of malaria. In many endemic areas, routine diagnosis is primarily performed microscopically, although rapid diagnostic tests (RDTs) provide a useful point-of-care tool. Most of the commercially available RDTs detect histidine-rich protein 2 (HRP2) of Plasmodium falciparum in the blood of infected individuals. Nonetheless, parasite isolates lacking the pfhrp2 gene are relatively frequent in some endemic regions, thereby hampering the diagnosis of malaria using HRP2-based RDTs. To track the efficacy of RDTs in areas of the Brazilian Amazon, we assessed pfhrp2 deletions in 132 P . falciparum samples collected from four malaria-endemic states in Brazil. Our findings show low to moderate levels of pfhrp2 deletion in different regions of the Brazilian Amazon. Overall, during the period covered by this study (2002-2020), we found that 10% of the P. falciparum isolates were characterized by a pfhrp2 deletion. Notably, however, the presence of pfhrp2 -negative isolates has not been translated into a reduction in RDT efficacy, which in part may be explained by the presence of polyclonal infections. A further important finding was the discrepancy in the proportion of pfhrp2 deletions detected using two assessed protocols (conventional PCR versus nested PCR), which reinforces the need to perform a carefully planned laboratory workflow to assess gene deletion. This is the first study to perform a comprehensive analysis of PfHRP2 sequence diversity in Brazilian isolates of P. falciparum . We identified 10 PfHRP2 sequence patterns, which were found to be exclusive of each of the assessed regions. Despite the small number of PfHRP2 sequences available from South America, we found that the PfHRP2 sequences identified in Brazil and neighboring French Guiana show similar sequence patterns. Our findings highlight the importance of continuously monitoring the occurrence and spread of parasites with pfrhp2 deletions, while also taking into account the limitations of PCR-based testing methods associated with accuracy and the complexity of infections., 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 © 2021 Costa, Mascarenhas, Martin, Fortini, Louzada, Pereira, Aguiar, Carvalho, de Brito, Fontes and de Sousa.)

The aim of the study was to evaluate the analgesic efficacy of tramadol compared to butorphanol administered locally in ventral surface of the hind paw of rats. Prospective, randomized experimental study; twenty-one adult male Wistar rats were selected. Heart rate (beats minute-1), respiratory rate (breaths minute-1), and paw withdrawal latency (onset of radiant heat until paw withdrawal/seconds) were measured prior (T0) and after (T5, T10, T15, T20) intraplantar injection with saline solution 0,9% (group S), butorphanol 1 mg kg-1 (group B), and tramadol 1 mg kg-1 (group T). Shapiro-Wilk normality test and Friedman test were used to analyze the data expressed by median and range. Statistical significance was set at p < 0.05. Statistical analysis of heart rate showed that there were significant differences between groups at different monitoring times. There were no significant differences in respiratory rate after intraplantar injection in any of the treatment groups. The paw withdrawal latency values at T5, T10, and T15 minutes after intraplantar injection in the group B were significantly higher compared to baseline value and to the values of the other groups. The paw withdrawal latency were no significant changes in the measurements of intragroup in S and T. Intraplantar administration of butorphanol provides a good analgesia and significantly increases paw withdrawal latency compared to tramadol. Intraplantar injection of butorphanol could be useful and safe and safe technique to achieve local analgesia for minor surgical procedures in rats., Competing Interests: The authors have declared that no competing interests exist.

The aim of this study was to identify possible predisposing anatomical factors associated with humeral fractures in birds of prey through comparison of specific anatomical features in different raptor species. An anatomical study of bone features in birds of prey was performed on 3 male subjects from 5 different species. The selected species included in this investigation were 3 diurnal species (the common buzzard [ Buteo buteo ], the peregrine falcon [ Falco peregrinus ], and the European honey-buzzard [ Pernis apivorus ]) and 2 nocturnal species (the barn owl [ Tyto alba ] and the tawny owl [ Strix aluco ]). Humeral bone samples were tomographically analyzed with a micro-macro-focus computed tomographic machine. Specific humeral anatomical points were selected (foramen pneumaticum and tuberculum dorsale for the proximal humerus; corpus humeri for the diaphyseal humerus; and above the condylus dorsalis for the distal humerus) to measure foramen pneumaticum diameter (in millimeters), cortical thickness (in millimeters), and trabeculae number and sizes (in millimeters). Apparent density, measured with the Hounsfield unit, was used to assess the degree of bone resistance. Statistical analysis was performed with a Spearman's correlation, and significance was set at P < .05. The differences among the observed bone volumes were highly significant ( P = .00). Trabeculae number and the humeral anatomical point measurements showed differences in all 5 avian species investigated. However, those differences were not significant, except at the condylus dorsalis ; in which, a significant interspecies difference ( P = .002) was recorded. Trabecular size, cortical thickness, bone density, and diameter of the foramen pneumaticum were all different in all raptor species; however, these variations were not significant. The study confirms the existence of humeral bone volume differences between diurnal and nocturnal species. Furthermore, the data suggest that the humeri of peregrine falcons and European honey-buzzards may be stronger than the humeri of common buzzards, tawny owls, and barn owls.

Background: The partial or complete cranial cruciate ligament rupture is a common skeletal disease affecting the stifle joint in dogs. The tibial plateau levelling osteotomy, performed with several synthesis systems, changed the approach to its treatment in dogs. The aim of this study was to compare two types of fixation implants, locking compression system and locking system, evaluating radiographically the progression of osteoarthritis of the stifle joint in dogs with complete cranial cruciate ligament deficiency treated surgically with tibial plateau levelling osteotomy. Moreover, we evaluated bone healing and lameness scores to show biomechanical effects by the implant used. Twenty-eight dogs, who met the inclusion criteria, were divided into two groups. Group A: 14 dogs treated using locking compression plates; Group B: 14 dogs treated using locking plates. Radiographic osteoarthritis scores were evaluated up to 1 year following tibial plateau levelling osteotomy. At each visit, animals were clinically and radiographically assessed. Each dog was evaluated before (T0) and after two (T2) and twelve (T12) months after the surgery. At T2 the stage of bone healing was evaluated. The clinical follow up was performed before the surgery and at 10, 15 and 20 days after the surgery, grading the lameness at walk and trot., Results: An increase in osteoarthritis score at T12 versus T0 in both groups was detected. A decrease of the lameness score was observed in Group A versus Group B. The healing score system at T2 showed a lower score in Group A versus Group B., Conclusions: The osteoarthritis score following tibial plateau levelling osteotomy did not differ when comparing the two different fixation systems. The locking compression system allowed a more rapid functional recovery of the limb and a quicker bone healing than the locking system. Locking compression system should be carefully considered for dogs subjected to tibial plateau levelling osteotomy surgery, because it may reduce the recovery time.

The trend in laparoscopy is to develop easy and rapid techniques associated with reduced intraoperative complications and decreased postoperative pain. The aim of this study was to compare combined laparoscopic ovariectomy (OIE) and laparoscopic-assisted incisional gastropexy (LAG) with combined laparoscopic OIE and total laparoscopic gastropexy (TLG) for surgical time, incidence of complications and postoperative pain. Twenty-eight female dogs were randomly assigned to the LAG group (n = 14) or the TLG group (n = 14). All laparoscopic procedures were performed using a three-port technique. The gastropexy was located 3 cm caudal to the 13th rib and 4 cm lateral to the rectus abdominis muscle. Surgical time (minutes [min]), intraoperative complications and postoperative complications were recorded. The Glasgow pain score (GPS) (short form) was calculated before surgery and at 1, 6, 12, 18 and 24 hr after extubation. Surgical time was significantly longer in the TLG group (48 ± 2 min) compared with the LAG group (39 ± 2 min). Minor postoperative complications occurred in both groups and included swelling (n = 2) and subcutaneous emphysema (n = 1). No significant differences regarding the GPS were recorded between groups. The GPS was significantly higher in both groups at 1 hr and 6 hr than before surgery. Two dogs in each group required rescue analgesia. Combined laparoscopic OIE and TLG require more time to perform than combined laparoscopic OIE and LAG. Neither procedure results in significant surgical complications. Postoperative pain for 24 hr was mild and comparable in both groups., (© 2020 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Objectives: The purpose of this study was to describe the metabolic and anthropometric profile of the Xikrin (Mebengôkre), an indigenous group., Methods: A total of 363 subjects (55.1% women) aged 18 years or older were evaluated. The variables analyzed were age, body weight and height, waist circumference, blood pressure, fasting blood glucose, triglyceride level, and total cholesterol level., Results: A high prevalence of obesity (36.5%) and central obesity (88.1%), mainly among women (46.9% and 96.2%, respectively), was found among the Xikrin (Mebengôkre). Impaired fasting glycemia and diabetes were found in 4.5% and 3.8% of adults, respectively. Twenty-one percent of adults had dyslipidemia and 9.3% had hypertension., Conclusions: The high prevalence of excess weight (overweight and obesity) and central obesity, especially in women, was the most significant finding of this study among the Xikrin (Mebengôkre)., (© 2019 Wiley Periodicals, Inc.)

This study assessed the effects of a combination of dexmedetomidine and butorphanol on the Schirmer tear test I (STT I) values in dogs. Ninety-eight dogs were sedated with an intramuscular injection of a combination of dexmedetomidine, 5 μg/kg body weight (BW), and butorphanol, 0.2 mg/kg BW. The effects of dexmedetomidine were reversed by administering atipamezole at the end of the procedure. The combination of dexmedetomidine and butorphanol significantly decreased tear production 15 minutes after sedation. The STT I values 15 minutes after reversal of dexmedetomidine with atipamezole were significantly higher than the STT I values 15 minutes after sedation but were significantly lower than the STT I values before sedation. Gender, weight, duration of sedation, right or left eye did not affect STT I values after sedation. It is recommended that dogs sedated with a combination of dexmedetomidine and butorphanol be treated with a tear substitute to combat decreased tear production.

Objective: To evaluate the anaesthetic effects of three different alfaxalone doses to induce anaesthesia in goldfish., Study Design: Prospective, randomized, clinical study., Animals: Thirty goldfish undergoing skin scraping, gill examination and stool collection., Methods: Each fish was transferred to an individual 4 L induction tank and randomly allocated into one of three groups (n = 10), in which alfaxalone was administered at concentrations of 6, 7 or 9 mg L -1 . The depth of anaesthesia was evaluated by approach reaction, equilibrium, opercular movement and reaction to tactile stimuli. Sedation, light anaesthesia, surgical anaesthesia and recovery times were recorded. Data were analyzed with analysis of variance. A p value <0.05 was considered significant., Results: Surgical anaesthesia was achieved in all fish. Goldfish induced with alfaxalone 7 and 9 mg L -1 showed a mild excitement phase. Time to sedation of the 6 mg L -1 dose (5.89 ± 0.40 minutes) was significantly longer compared to the 7 mg L -1 (3.97 ± 0.40 minutes) and 9 mg L -1 doses (3.94 ± 0.40 minutes). Times to light anaesthesia and surgical anaesthesia of the 9 mg L -1 dose (7.65 ± 1.04 and 9.60 ± 1.84 minutes, respectively) were significantly faster compared with those of the 6 mg L -1 dose (13.79 ± 1.04 and 19.75 ± 1.84 minutes, respectively) and the 7 mg L -1 dose (13.55 ± 1.04 and 21.24 ± 1.84 minutes, respectively). No significant differences were recorded in recovery time. Cessation of opercular movement was recorded in two fish induced with 7 mg L -1 and in two induced with 9 mg L -1 . No mortality occurred., Conclusions: and clinical relevance Alfaxalone is a reliable agent for immersion anaesthesia in goldfish. Immersion in water containing 6 mg alfaxalone L -1 provided smooth induction of anaesthesia, and no obvious side effects were encountered. Higher doses shortened induction time and caused respiratory depression and excitatory movements., (Copyright © 2018 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

Objective: The aim of this study was to evaluate the reliability of different dynamometric variables of the pelvic floor muscles (PFM) in healthy women during different periods of menstrual cycle., Materials and Methods: Vaginal dynamometric equipment was developed by the authors and its reproducibility was tested. The PFM contractions of 20 healthy women were collected by two independent examiners over three consecutive weeks, always on the same day, with a seven-day interval between readings, starting from the first day after the end of the menstrual period. For the measurements, the branch of the dynamometer was positioned first on the sagittal plane and then on the frontal plane. Baseline, peak time, maximum PFM strength, impulse contraction, and average contraction force were calculated. Reproducibility was tested using the intra-class correlation coefficient (ICC) and standard error of measurement. Repeated-measures ANOVA was used to compare the data from different days., Results: For intra-day and inter-day reliability between examiners, all the parameters collected on the sagittal plane presented good and excellent reproducibility (ICC 2,1  = 0.60 to 0.98), whereas reproducibility on the frontal plane was respectively poor and excellent (ICC 2,1  = 0.23 to 0.97). The ANOVA revealed significant differences between sessions only for the impulse of contraction for the sagittal (P = 0.005) and frontal (P = 0.03) planes., Conclusions: Time and contraction force parameters of the PFM are not influenced by hormonal alterations that occur during the menstrual cycle. The impulse of contraction was the only variable to demonstrate a significant difference between the first and second week of the data collection protocol. The baseline, maximum strength value, impulse of contraction, and average contraction force variables presented good to excellent reproducibility and can be safely used as a method of PFM evaluation., (© 2018 Wiley Periodicals, Inc.)

Background: The resistance of Plasmodium vivax to chloroquine has become an obstacle to control strategies based on the use of anti-malarials. The current study investigated the association between P. vivax CQ-resistance in vivo with copy number variation and mutations in the promoter region in pvcrt-o and pvmdr1 genes., Methods: The study included patients with P. vivax that received supervised treatment with chloroquine and primaquine. Recurrences were actively recorded during this period., Results: Among the 60 patients with P. vivax, 25 were CQ-resistant and 35 CQ-susceptible. A frequency of 7.1% of multi-copy pvcrt-o was observed in CQ-susceptible samples and 7.7% in CQ-resistant at D0 (P > 0.05) and 33.3% in CQ-resistant at DR (P < 0.05). For pvmdr1, 10.7% of the CQ-susceptible samples presented multiple copies compared to 11.1% in CQ-resistant at D0 and 0.0% in CQ-resistant at DR (P > 0.05). A deletion of 19 bp was found in 11/23 (47.6%) of the patients with CQ-susceptible P. vivax and 3/10 (23.1%) of the samples with in CQRPv at D0. At day DR, 55.5% of the samples with CQRPv had the 19 bp deletion. For the pvmdr-1 gene, was no variation in the analysed gene compared to the P. vivax reference Sal-1., Conclusions: This was the first study with 42-day clinical follow-up to evaluate the variation of the number of copies and polymorphisms in the promoter region of the pvcrt-o and pvmdr1 genes in relation to treatment outcomes. Significantly higher frequency of multi-copy pvcrt-o was found in CQRPv samples at DR compared to CQ-susceptible, indicating parasite selection of this genotype after CQ treatment and its association with CQ-resistance in vivo.

[This corrects the article DOI: 10.1371/journal.pone.0160172.].

Background: Parasite resistance to anti-malarials represents a great obstacle for malaria elimination. The majority of studies have investigated the association between single-nucleotide polymorphisms (SNPs) and drug resistance; however, it is becoming clear that the copy number variation (CNV) is also associated with this parasite phenotype. To provide a baseline for molecular surveillance of anti-malarial drug resistance in the Brazilian Amazon, the present study characterized the genetic profile of both markers in the most common genes associated with drug resistance in Plasmodium falciparum and Plasmodium vivax isolates. Additionally, these data were compared to data published elsewhere applying a systematic review of the literature published over a 20-year time period., Methods: The genomic DNA of 67 patients infected by P. falciparum and P. vivax from three Brazilian States was obtained between 2002 and 2012. CNV in P. falciparum multidrug resistance gene-1 (pfmdr1), GTP cyclohydrolase 1 (pfgch1) and P. vivax multidrug resistance gene-1 (pvmdr1) were assessed by real-time PCR assays. SNPs in the pfmdr1 and pfcrt genes were assessed by PCR-RFLP. A literature search for studies that analysed CNP in the same genes of P. falciparum and P. vivax was conducted between May 2014 and March 2017 across four databases., Results: All analysed samples of P. falciparum carried only one copy of pfmdr1 or pfgch1. Although the pfcrt K76T polymorphism, a determinant of CQ resistance, was present in all samples genotyped, the pfmdr1 N86Y was absent. For P. vivax isolates, an amplification rate of 20% was found for the pvmdr1 gene. The results of the study are in agreement with the low amplification rates for pfmdr1 gene evidenced in the Americas and Africa, while higher rates have been described in Southeast Asia. For P. vivax, very low rates of amplification for pvmdr1 have been described worldwide, with exceptions in French Guiana, Cambodia, Thailand and Brazil., Conclusions: The present study was the first to evaluate gch1 CNV in P. falciparum isolates from Brazil, showing an absence of amplification of this gene more than 20 years after the withdrawal of the Brazilian antifolates therapeutic scheme. Furthermore, the rate of pvmdr1 amplification was significantly higher than that previously reported for isolates circulating in Northern Brazil.

We used physiological parameters and the duration and quality of anesthesia to compare the effects of two ambient temperatures and of the duration of pre-immobilization confinement on the chemical immobilization of fallow deer. We divided 45 free-ranging fallow deer ( Dama dama ) into two groups: Group A were deer captured in winter (average 12 C), using 1 mg/kg of xylazine and 1 mg/kg of tiletamine-zolazepam; and Group B were deer captured in spring (average 24 C), using 2 mg/kg of xylazine and 1.5 mg/kg of tiletamine-zolazepam, after being confined in a pen. We observed lower mean respiratory rate and oxygen saturation in Group B. In contrast, the mean body temperature and the mean blood lactate concentration were significantly higher in Group B, and quality of anesthesia was better in Group A. Mean induction time (time to achieve recumbency) and durations of recumbency were the same in Groups A and B: approximately 8 and 50 min, respectively. Despite the lower drug dosage, better sedation was obtained in Group A than in group B. The time of year, most likely associated with differences in ambient temperature and in confinement, influenced the recommended dosage for xylazine and tiletamine-zolazepam in fallow deer. As all the animals were sound, we concluded that the only factors that influenced the outcome of the present study were the ambient temperature and the level of stress caused by confinement in the pen.

Antonio-Santos, J, Ferreira, DJS, Gomes Costa, GL, Matos, RJB, Toscano, AE, Manhães-de-Castro, R, and Leandro, CG. Resistance training alters the proportion of skeletal muscle fibers but not brain neurotrophic factors in young adult rats. J Strength Cond Res 30(12): 3531-3538, 2016-Resistance training (RT) is related to improved muscular strength and power output. Different programs of RT for rats have been developed, but peripheral and central response has not been evaluated directly in the same animal. To test the hypothesis that RT induces central and peripheral adaptations, this study evaluated the effects of a RT on the performance of a weekly maximum overload test, fiber-type typology, and brain neurotrophic factors in young adult rats. Thirty-one male Wistar rats (65 ± 5 days) were divided in 2 groups: nontrained (NT, n = 13) and trained (T, n = 18). Trained group was submitted to a program of RT ladder climbing, gradually added mass, 5 days per week during 8 weeks at 80% of individual maximum overload. This test was weekly performed to adjust the individual load throughout the weeks for both groups. After 48 hours from the last session of exercise, soleus and extensor digital longus (EDL) muscles were removed for myofibrillar ATPase staining analysis. Spinal cord, motor cortex, and cerebellum were removed for RT-PCR analysis of BDNF and insulin-like growth factor-1 (IGF-1) gene expression. In EDL muscle, T animals showed an increase in the proportion of type IIb fibers and a reduction of type IIa fibers. Insulin-like growth factor-1 gene expression was reduced in the cerebellum of T animals (NT: 1.025 ± 0.12; T: 0.57 ± 0.11). Our data showed that 8 weeks of RT were enough to increase maximum overload capacity and the proportion of glycolytic muscle fibers, but there were no associations with the expression of growth neurotrophic factors.

Although Plasmodium vivax relapses are classically associated with hypnozoite activation, it has been proposed that a proportion of these cases are due to primaquine (PQ) treatment failure caused by polymorphisms in cytochrome P-450 2D6 (CYP2D6). Here, we present evidence that CYP2D6 polymorphisms are implicated in PQ failure, which was reinforced by findings in genetically similar parasites, and may explain a number of vivax relapses. Using a computational approach, these polymorphisms were predicted to affect the activity of CYP2D6 through changes in the structural stability that could lead to disruption of the PQ-enzyme interactions. Furthermore, because PQ is co-administered with chloroquine (CQ), we investigated whether CQ-impaired metabolism by cytochrome P-450 2C8 (CYP2C8) could also contribute to vivax recurrences. Our results show that CYP2C8-mutated patients frequently relapsed early (<42 days) and had a higher proportion of genetically similar parasites, suggesting the possibility of recrudescence due to CQ therapeutic failure. These results highlight the importance of pharmacogenetic studies as a tool to monitor the efficacy of antimalarial therapy.

A new halimane diterpene was isolated from Vellozia kolbekii Alves (Velloziaceae) and identified as (5R,8R,9S,13R)-halim-1,10-ene-15,16-diol (1). It showed cytotoxicity against three human cancer cell lines, SF-295 (glioblastoma), MDA-MB-435 (melanoma), and HCT-8 (colon adenocarcinoma). In the mechanism of cytotoxic action, halimane 1 interferes in two major phases of the cell cycle: in S phase, in which DNA synthesis occurs and where it is very sensitive to damage, and G2M phase which is the phase of preparation for mitosis and mitosis itself, showing apoptosis-inducing properties. Antimicrobial activity towards Gram-positive and Gram-negative bacteria was studied and, against Bacillus cereus, B. subtilis, Escherichia coli, and Pseudomonas aeruginosa, a MIC value of 0.025 μM was observed for halimane 1, which is more active than the positive control chloramphenicol., (Copyright © 2015 Verlag Helvetica Chimica Acta AG, Zürich.)

Objective: To evaluate the efficacy and safety of the surgical technique called the "parachute technique, as applied to adult patients who suffered displaced and unstable two or three-part fractures of the proximal humerus, through the clinical outcomes., Methods: Between January 1995 and June 2006, 59 adult patients with displaced and unstable two or three-part fractures underwent operations performed by the Shoulder and Elbow Group of the Orthopedics and Traumatology Service of Hospital do Servidor Público Estadual de São Paulo using the "parachute technique. This method consists of an intramedullary tension band and extramedullary fixation in a figure-of-eight to join the fragments of the fracture, using a 6.5-millimeter spongy screw with partial threading, a washer and two non-absorbable wires, thereby producing stable synthesis with minimal aggression to the surrounding soft tissue and not requiring any subsequent removal of material. The final shape of this synthesis is reminiscent of the shape of an open parachute. The patients had a minimum postoperative follow-up of six months. For the diagnosis, trauma series radiographic views of the shoulder were produced. The fractures were classified in accordance with the system proposed by Neer. We used the scale of the University of California, Los Angeles (UCLA), to evaluate the results., Results: The "parachute technique" produced good results in 47% and excellent results in 26% of the cases, according to the UCLA scores., Conclusion: The "parachute technique" is a safe and effective treatment for displaced and unstable two or three-part fractures of the proximal humerus.

The influenza virus causes acute respiratory infections, leading to high morbidity and mortality in groups of patients at higher risk. Antiviral drugs represent the first line of defense against influenza, both for seasonal infections and pandemic outbreaks. Two main classes of drugs against influenza are in clinical use: M2-channel blockers and neuraminidase inhibitors. Nevertheless, because influenza strains that are resistant to these antivirals have been described, the search for novel compounds with different mechanisms of action is necessary. Here, we investigated the anti-influenza activity of a fungi-derived natural product, aureonitol. This compound inhibited influenza A and B virus replication. This compound was more effective against influenza A(H3N2), with an EC50 of 100 nM. Aureonitol cytoxicity was also very low, with a CC50 value of 1426 μM. Aureonitol inhibited influenza hemagglutination and, consequently, significantly impaired virus adsorption. Molecular modeling studies revealed that aureonitol docked in the sialic acid binding site of hemagglutinin, forming hydrogen bonds with highly conserved residues. Altogether, our results indicate that the chemical structure of aureonitol is promising for future anti-influenza drug design.

Objective: To evaluate the anti-nociceptive and sedative effects of slow intravenous (IV) injection of tramadol, romifidine, or a combination of both drugs in ponies., Study Design: Within-subject blinded., Animals: Twenty ponies (seven male, 13 female, weighing mean ± SD 268.0 ± 128 kg)., Methods: On separate occasions, each pony received one of the following three treatments IV; romifidine 50 μg kg(-) (R) tramadol 3 mg kg(-1) given over 15 minutes (T) or tramadol 3 mg kg(-1) followed by romifidine 50 μg kg(-1) (RT). Physiologic parameters and caecal borborygmi (CB) were measured and sedation and response to electrical stimulation of the coronary band assessed before and up to 120 minutes following drugs administration. Results were analyzed using the Friedman's test and 2 way anova as relevant., Results: When compared to baseline, heart (HR, beats minute(-1) ) and respiratory rates (fR , breaths minute(-1) ) increased with treatment T (highest mean ± SD, HR 43 ± 1; fR 33 ± 2) and decreased with R (lowest HR 29 ± 1 and fR 10 ± 4) and RT (lowest HR 32 ± 1 and fR 9 ± 3). There were no changes in other measured physiological variables. The height of head from the ground was lower following treatments R and TR than T. There was slight ataxia with all three treatments. No excitatory behavioural effects were observed. The response to electrical stimulation was reduced for a prolonged period relative to baseline following all three treatments, the effect being significantly greatest with treatment RT., Conclusion: Tramadol combined with romifidine at the stated doses proved an effective sedative and anti-nociceptive combination in ponies, with no unacceptable behavioural or physiologic side effects., Clinical Relevance: Slow controlled administration of tramadol should reduce the occurrence of adverse behavioural side effects., (© 2014 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia.)

Background: The efficacy and risks of oral anticoagulation are largely associated with maintaining the quality of anticoagulation control. Nevertheless, few studies have addressed which factors, if any, are associated with this control., Objective: This study aimed to identify predictors of high-quality oral anticoagulation., Methods: A prospective observational study enrolled all adult patients on intended long-term oral anticoagulation attending a public anticoagulation clinic. Patients with high-quality anticoagulation, defined as percentage of time in therapeutic international normalized ratio (INR) range (TTR) ≥66%, were compared with those with poor anticoagulation control (TTR <66%). Measures included cognitive, psychological, and relevant behavioral factors, in addition to traditionally implicated ones, such as age, comorbidity, and concurrent medications., Results: Participation was requested from all 233 patients followed up at the anticoagulation clinic. Eighty-six did not meet the inclusion criteria (49 due to intended anticoagulation duration <90 days, 37 due to the need for a caregiver responsible for medications). A total of 147 patients were enrolled, of whom 13 (8.8%) were lost to follow-up. Therefore, data were analyzed from 134 patients (mean [SD] age, 55 [14.2] years [range, 19-87 years]), who were followed up for a mean (SD) duration of 272 (87) days. The total mean TTR was 64.7%, which is comparable to values achieved in clinical trials. The good-control group had 61 patients (45.5%) (mean TTR, 77.7% [8.5%]) and the poor-control group had 73 patients (54.5%) (mean TTR, 50.4 [11.7%]). On multivariate logistic regression analysis, high-quality anticoagulation was independently associated with regular vitamin K intake, expressed by its variability in daily dosage (odds ratio [OR] = 0.79; 95% CI, 0.64-0.98); male sex (OR = 2.41; 95% CI, 1.06-5.49); duration of anticoagulation treatment >2 months (OR = 3.23; 95% CI, 1.25-8.36); presence of family support (OR = 3.32; 95% CI, 1.16-9.48); functional and cognitive ability to take medications as prescribed, defined as good medication management capacity (MMC; as assed using the Drug Regimen Unassisted Grading Scale) (OR = 4.18; 95% CI, 1.63-10.68); and no regular use of alcohol (OR = 8.59; 95% CI, 1.45-51.09)., Conclusions: The data suggest that independent predictors of high-quality oral anticoagulation included regular vitamin K intake, male sex, duration of anticoagulation treatment >2 months, presence of family support, good MMC, and no regular alcohol use. These findings may help clinicians to decide whether to start anticoagulation in intermediate-risk patients, to identify patients who will require closer attention on their anticoagulation management, and to direct their efforts to improve the quality of oral anticoagulation., (Copyright © 2012 Elsevier HS Journals, Inc. All rights reserved.)

Penicillium corylophilum isolated from mosquitoes was cultivated in liquid media leading to the first reported identification of citrinin (1a) as one metabolic component of this fungus. The produced amount of 1a indicated this compound as the most abundant secondary metabolite of this fungus. Stressing the culture of P. corylophilum with the presence of the antagonistic fungus Beauveria bassiana increased in 65% the production of 1a. Similar results were obtained with the presence of other fungi in the culture media, indicating that citrinin is the main defence metabolite of P. corylophilum. In agreement with this conclusion, citrinin showed a reasonable fungicidal activity against Colletotrichum gloeosporioides and B. bassiana.

Endostatin is a potent antiangiogenic protein derived from the noncollagenous domain 1 (NC1) of collagen XVIII. The mechanism by which endostatin exerts its antiangiogenic effect is still incompletely understood. It has been shown that the 27 amino acid N-terminal fragment of murine endostatin has antitumor, antimigration, and antipermeability activities comparable to the full soluble protein. To understand how this peptide can exert such elaborate function, we performed structural analysis using molecular dynamics to evaluate the behavior of this fragment in aqueous environment. Here, we show that the N-terminal peptide of murine endostatin is able to assume a well-defined structure, folding into a zinc-dependent β-hairpin conformation. Analyzing the folding mechanism, we were able to understand why the N-terminal peptide of human endostatin with the same length failed to acquire a stable conformation. Conversely, we were able to predict the successful folding of the R4Q mutant and of a shorter form of the human peptide with 25 residues. Finally, we show that the β-hairpin conformation assumed by the zinc-bound peptide of murine endostatin has a high structural similarity with fragments of another family of angiogenesis inhibitors: the integrin-binding portion of the NC1 domain of collagen IV. Indeed, our docking simulations show that arresten, canstatin, and the endostatin peptide bind to the same spot of αVβ3 integrin, suggesting similar interactions via a common binding site on this receptor., (Copyright © 2011 Wiley-Liss, Inc.)

Background: despite the overwhelming evidence of its effectiveness, oral anticoagulation continues to be underused in the elderly, presumably due to physicians' misconceptions when estimating bleeding risk and ability to comply with treatment., Objective: to investigate the quality of anticoagulation control among deprived elderly and younger patients., Design: prospective observational study., Setting: a public anticoagulation clinic in a developing country., Subjects: all adult patients on intended long-term (>90 days) oral anticoagulation. We studied 171 patients (79 elderly and 92 non-elderly) with a mean follow-up of 273 ± 84.9 days., Methods: the main outcome measure was the quality of anticoagulation management as measured by the time in therapeutic (TTR) international normalised ratio (INR) range. Elderly patients (≥60 years) were compared with younger patients with respect to the educational level and co-morbidities., Results: the mean number of years of formal education was 4.37 ± 3.2 years. The mean TTR was 62.50 ± 17.9% in non-elderly and 62.10 ± 16.6% in elderly (P = 0.862) subjects, despite the higher prevalence of co-morbidities in the latter group: heart failure (46.3 versus 28.6%, P = 0.042), diabetes mellitus (22.8 versus 8.7%, P = 0.011), renal failure (estimated glomerular filtration rate <50 ml/min: 38.0 versus 7.1%, P < 0.001) and polypharmacy (84.8 versus 58.7%, P < 0.001). Fifty elderly and 84 non-elderly subjects require little or no assistance in taking medications. Among them, the elderly had lower educational levels (3.42 ± 2.5 versus 5.55 ± 3.4 years of formal education, P < 0.001) and higher rates of cognitive impairment (34.0 versus 13.1%, P = 0.004), but a similar mean TTR (62.46 ± 16.1 versus 63.02 ± 17.8%, P = 0.856). The oldest (≥75 years) patients did as well as those aged ≤50 years (mean TTR: 62.54 ± 16.0 versus 62.23 ± 16.4%, respectively, P = 0.98)., Conclusions: good-quality management of oral anticoagulation is achievable in deprived populations attending an anticoagulation clinic. Elderly patients may experience similar quality of anticoagulation despite having higher levels of co-morbidities and polypharmacy. These results add evidence to the safety of such therapeutic interventions in the elderly.

Scleredema of Buschke (SB) is a rare disorder of connective tissue, characterized by hardening of the skin. Of unknown etiology, it may be associated with viral or bacterial infections, hematological alterations and diabetes mellitus (DM). In half of the reported cases of SB, there is association with DM. The most common characteristics among these patients are: being male adults, having long-term DM, bad metabolic control and presence of specific complications of DM. The histopathological examination shows dermal thickening, with widened collagen fibers separated by non-colored spots, which correspond to mucopolysaccharides deposits. The clinical consequences are: decrease in motility of the shoulders and an impairment of respiratory function. Several treatments are suggested in the literature, but with inconstant results. A case of SB is reported in a type 2 diabetes patient.

Cancer is driven by mutation. Worldwide, tobacco smoking is the principal lifestyle exposure that causes cancer, exerting carcinogenicity through >60 chemicals that bind and mutate DNA. Using massively parallel sequencing technology, we sequenced a small-cell lung cancer cell line, NCI-H209, to explore the mutational burden associated with tobacco smoking. A total of 22,910 somatic substitutions were identified, including 134 in coding exons. Multiple mutation signatures testify to the cocktail of carcinogens in tobacco smoke and their proclivities for particular bases and surrounding sequence context. Effects of transcription-coupled repair and a second, more general, expression-linked repair pathway were evident. We identified a tandem duplication that duplicates exons 3-8 of CHD7 in frame, and another two lines carrying PVT1-CHD7 fusion genes, indicating that CHD7 may be recurrently rearranged in this disease. These findings illustrate the potential for next-generation sequencing to provide unprecedented insights into mutational processes, cellular repair pathways and gene networks associated with cancer.

We describe the genome sequencing of an anonymous individual of African origin using a novel ligation-based sequencing assay that enables a unique form of error correction that improves the raw accuracy of the aligned reads to >99.9%, allowing us to accurately call SNPs with as few as two reads per allele. We collected several billion mate-paired reads yielding approximately 18x haploid coverage of aligned sequence and close to 300x clone coverage. Over 98% of the reference genome is covered with at least one uniquely placed read, and 99.65% is spanned by at least one uniquely placed mate-paired clone. We identify over 3.8 million SNPs, 19% of which are novel. Mate-paired data are used to physically resolve haplotype phases of nearly two-thirds of the genotypes obtained and produce phased segments of up to 215 kb. We detect 226,529 intra-read indels, 5590 indels between mate-paired reads, 91 inversions, and four gene fusions. We use a novel approach for detecting indels between mate-paired reads that are smaller than the standard deviation of the insert size of the library and discover deletions in common with those detected with our intra-read approach. Dozens of mutations previously described in OMIM and hundreds of nonsynonymous single-nucleotide and structural variants in genes previously implicated in disease are identified in this individual. There is more genetic variation in the human genome still to be uncovered, and we provide guidance for future surveys in populations and cancer biopsies.

Objective: The elderly constitute a population with their own features and frequent admissions in intensive care units. This study has the objective to evaluate the ability to predict the survival of these patients through the APACHE II, UNICAMP II, SAPS II and SAPS 3 indexes, global and Central America/South equations., Methods: Elderly patients admitted from 01/01/2006 to 12/3/2006, defined as age > 60 years, were included in this study. Those who were readmitted were excluded. The rate of lethality standardized, calibration and discrimination for each index in the remaining patients were analysed. The outcome were death or hospital discharge., Results: Three hundred eighty six elderly patients were included in this study, being 36 excluded by readmission, remaining 350 for analysis. The rate of lethality standardized came near to the unit in all indexes, except the SAPS II (TLP=1.5455) which underestimated the lethality. The calibration, via Hosmer-Lemeshow tests was inadequate (p < 0.05), except for the UNICAMP II (p > 0.5). On the calibration curve, the models have distanced themselves from the pattern line. All of them presented an excellent discrimination via receiver operating characteristics curves (> 0.8)., Conclusions: In the studied population, the models presented an excellent discrimination and inadequate calibration. SAPS II underestimated the lethality.

Ctenocephalides felis felis is a flea species that parasitizes dogs and cats. The current study verified the in vitro virulence of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana to eggs and adults of C. felis felis. The fungal isolates were cultivated on rice grains in plastic bags. Conidial suspensions were then prepared by immersing the grains in Tween 80 aqueous solution (0.01%). Eggs and adults of C. felis felis were obtained from a laboratory colony. Experiments were composed of one control and four treatment groups (10(5), 10(6), 10(7), and 10(8) conidia mL(-1)). Both insect pathogens inhibited egg hatchability and caused adult mortality. Among isolates used in this study, M. anisopliae (isolate E9) was the most successful at inhibiting hatching, while B. bassiana (isolate 986) was the most powerful for causing mortality in adults. The results indicate that these entomopathogenic fungi have promising potential for use in controlling both C. felis felis egg and adult stages.

Styrene-7,8-oxide (SO), a chemical compound widely used in industrial applications, is a potential hazard for humans, particularly in occupational settings. Neurobehavioral changes are consistently observed in occupationally exposed individuals and alterations of neurotransmitters associated with neuronal loss have been reported in animal models. Although the toxic effects of styrene have been extensively documented, the molecular mechanisms responsible for SO-induced neurotoxicity are still unclear. A possible dopamine-mediated effect of styrene neurotoxicity has been previously demonstrated, since styrene oxide alters dopamine neurotransmission in the brain. Thus, the present study hypothesizes that styrene neurotoxicity may involve synaptic contacts. Primary striatal neurons were exposed to styrene oxide at different concentrations (0.1-1 mM) for different time periods (8, 16, and 24 h) to evaluate the dose able to induce synaptic impairments. The expression of proteins crucial for synaptic transmission such as Synapsin, Synaptophysin, and RAC-1 were considered. The levels of Synaptophysin and RAC-1 decreased in a dose-dependent manner. Accordingly, morphological alterations, observed at the ultrastructural level, primarily involved the pre-synaptic compartment. In SO-exposed cultures, the biochemical cascade of caspases was activated affecting the cytoskeleton components as their target. Thus the impairments in synaptic contacts observed in SO-exposed cultures might reflect a primarily morphological alteration of neuronal cytoskeleton. In addition, our data support the hypothesis developed by previous authors of reactive oxygen species (ROS) initiating events of SO cytotoxicity.

This study was carried out with the objective of evaluating the efficacy of a 2,6-dichlorophenol (2,6-DCP) lure to control Dermacentor nitens (Acari: Ixodidae). Slow-release formulations of the pheromone formulated with and without cypermethrin were prepared. Olfactometer bioassays were used to define the best dose of the pheromone and to evaluate the effect of cypermethrin with 2,6-DCP attractiveness. Sexually active males were released 15 cm from 2 cmx1 cm pieces of polypropylene treated with different odors: 2,6-DCP in a liposphere system (1.5, 30 and 300 microg--without cypermethrin and 30 microg--with cypermethrin) and 2,6-DCP in hexane (30 microg). The tests were conducted 7 and 15 days after the preparation of the odor sources. The percentages of males that oriented, or showed directional movement toward the stimulus, and their tracks were recorded. Lures (10 cmx2 cm pieces of polypropylene) treated with the best dose of the liposphere formulation (30 microg) were prepared. The lures were evaluated in horses that had been artificially infested with D. nitens. The horses' ears were infested with 3000 D. nitens larvae per ear, once weekly for 4 weeks. The animals were divided into three groups: control, 2,6-DCP and 2,6-DCP+cypermethrin. On day 0, the lures of their respective treatments were attached to the horses' napes. From days 6 to 20 after attachment, female ticks of 4 mm or over in length were counted on the ears, every 2 or 3 days. Olfactometer analysis showed higher orientation to 30 microg dose and more prolonged release of the pheromone in the liposphere formulation than in hexane; cypermethrin did not interfere with the attractive effect of the pheromone. The lures were efficient in the first 10 days after attachment, when the mean number of females was higher in the control group (24.9) than in 2,6-DCP and cypermethrin (5.4), and 2,6-DCP (9.2) groups. After that period the number of females was similar in the control and treated groups. These results indicate that 2,6-DCP lures used in a liposphere formulation can control D. nitens for at least 10 days by preventing its copulation. However, further evaluation of longer-term pheromone release under natural conditions is needed in order to validate this kind of control. In addition, the use of extra lures on the horse's tail may help to control populations on the hindquarters.

Fifty isolates of Beauveria bassiana (Balsamo) Vuillemin, 1912 (Ascomycota: Clavicipitaceae) were analyzed by morphology, for their pathogenic potential to Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) larvae, and by Random Amplified Polymorphic DNA-Polymerase Chain Reaction technique. Morphological analysis demonstrated that isolates present characteristics compatible to those described for B. bassiana in the literature. Virulence test demonstrated that all isolates present lethal effect on larvae and that the lethal concentration varies among isolates. The most virulent isolate was the only one obtained from human infection, which was also the only isolate presenting synnemata. The study on genetic variability among the isolates allowed the identification of 23 electrophoretic profiles. The established groupings suggest that most of the isolates obtained from B. microplus of the same locality present low genetic variation. In this way, the data in the present study will contribute to a meticulous characterization of these B. bassiana isolates.

L-asparaginase production was investigated in the filamentous fungi Aspergillus tamarii and Aspergillus terreus. The fungi were cultivated in medium containing different nitrogen sources. A. terreus showed the highest L-asparaginase (activity) production level (58 U/L) when cultivated in a 2% proline medium. Both fungi presented the lowest level of L-asparaginase production in the presence of glutamine and urea as nitrogen sources. These results suggest that L-asparaginase production by of filamentous fungi is under nitrogen regulation.

The purpose of this work was to evaluate the in vitro virulence of three isolates of Metarhizium anisopliae var. anisopliae to eggs and larvae of the tick Boophilus microplus. The fungus tested was isolated from engorged females of B. microplus collected in the field, and identified as Ma01, Ma02 e Ma04. These isolates were evaluated by immersion of eggs and larvae in suspension with different conidial concentrations: 10(5), 10(6), 10(7) e 10(8) conidia/ml. In each isolate there was a treatment group for each spore concentration and a control group with 10 repetitions. It was observed in the treated egg groups that there was a hatching percentage that was much less than that observed in the control groups. This was in inverse proportion to the conidia concentration/ml. Larval bioassays of all the tested isolates resulted in a high mortality of larvae in direct proportion to the spore concentration/ml, 10 days after the conidia suspensions were inoculated. To consolidate the infection, the fungus used in bioassays was re-isolated., (Copyright 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Two strains, Penicillium corylophilum and Aspergillus giganteus, of the most frequent species found in a survey of triatomines, were used for bioassays in the second and fourth nymphs stage of Triatoma infestans and Panstrongylus megistus. Two procedures, bite and pulverization, were used and compared. A. giganteus was most effective, causing mortality in at least 50% of the nymphs of the two species tested with exception of the nymphs of the fourth stage of P. megistus. Variation in entomopathogenic capacity of the fungal species were observed in the experiments. The two procedures used proved effective.

The purpose of this work was to evaluate the in vitro virulence of three isolates of Beauveria bassiana to eggs and larvae of the tick Boophilus microplus. The fungus tested were isolated from engorged females of B. microplus collected on the field, and identified as Bb28, Bb29 and Bb30. These isolates were evaluated by immersion of eggs and larvae in suspensions with different conidial concentrations: 10(8), 10(7), 10(6) and 10(5) conidia/ml and compared to the control groups. In the treated eggs, there was a percentage much smaller of hatching than that observed in the controls. The egg hatch was inversely proportional to the conidial concentration. In larval bioassays, all isolates resulted in a higher mortality of larvae compared to the control according to the conidial concentrations/ml, 10 days after treatment inoculation.

The objective of this study was to isolate and identify fungal species found in natural association with adults of Musca domestica. The adult insects were collected from two natural breeding grounds: hog pens and an urban sanitary landfill. The isolated fungi were identified as: Aspergillus flavus (23.8%), A. niger var. niger (14.4%), Penicillium corylophilum (21.4%), P. fellutanum (11.9%), Cladosporium cladosporoides (4.7%), Fusarium sp. (4.7%), Alternaria alternata (11.9%), Curvularia brachyspora (2.4%), Mycelia sterilia (2.4%) and the Mucorales order (2.4%).