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The potential of bitter orange peel powder (BOPP) as a nutritional strategy for fish was investigated in Nile tilapia. A total of 120 juveniles with an average initial weight of 9.8 ± 0.7 g were divided into four groups, replicated three times, resulting in 12 experimental units (60 L each) at a stocking density of 1.63 g of fish per liter. Productive parameters, whole-body composition, blood biochemistry, erythroid morphometry, intestinal histology, and heat tolerance were assessed in the juveniles subjected to one of the following treatments: non-supplemented basal diet (control group); basal diet with BOPP at 10 g/kg (BOPP10 group); basal diet with BOPP at 20 g/kg (BOPP20 group); and basal diet with BOPP at 40 g/kg (BOPP40 group). The BOPP additive had a positive influence on Nile tilapia growth, as final weight and weight gain were greater in all BOPP-treated fish, despite the reduction in crude protein in BOPP10 and BOPP20 groups. Fish receiving BOPP40 had an increase in total lipids and showed the highest levels of triglycerides and total cholesterol. Villi development was greater in the tilapia given BOPP10. It may be concluded that BOPP presented the most promising results for Nile tilapia juveniles when used at 10 g/kg diet. Regarding the erythroid morphometry, there was a general increase in nuclear and cytoplasmic areas in BOPP-fed tilapia; this seems to be the first report on the direct impact of the inclusion of functional additives in fish diet upon such parameters. As concerns the thermal tolerance evaluated at the end of the feeding trial, no differences were registered among the experimental groups. Thus, BOPP represents a feasible alternative ingredient to be explored in fish nutrition, since orange peel is a natural low-cost source of essential nutrients and valuable bioactive compounds., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Sarcocystis spp. are protozoan parasites that form cysts in the organs and musculature of various animal species. The species Sarcocystis miescheriana and Sarcocystis suihominis are pathogenic to pigs and wild boars (Sus scrofa), acting as intermediate hosts, while humans are the definitive host for S. suihominis. To date, there have been no reports of the identification of these coccidian species in Sus scrofa in Brazil. Therefore, in this study, we conducted the first molecular identification of Sarcocystis species using PCR-RFLP and sequencing. A total of 210 samples were analyzed, of this total, 67 tested positive for Sarcocystis spp., representing 31.9% of the total samples assessed. Out of the total positive samples, 55 (82.1%) were identified as S. miescheriana and 8 (11.9%) as S. suihominis, a zoonotic species. Additionally, other species related to bovines, such as S. cruzi and zoonotic S. hominis, were detected in 3.0% of the samples, serving as contaminants in the pork products. The presence of S. suihominis in swine and wild boar samples is concerning due to the zoonotic risk and potential environmental contamination, as humans act as definitive hosts, also for the presence of S. hominis as a bovine contaminant in pork sausages. Furthermore, we confirmed the efficacy of the PCR-RFLP technique as a reliable tool for the identification of Sarcocystis species, demonstrating its potential use in laboratories for molecular diagnosis and rapid identification of these parasites, aiming to protect public health and ensure food safety., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Trypanosoma evansi is a flagellate protozoan that infects a wide range of hosts, especially horses. Clinically, the infection is characterized by rapid weight loss, anemia and mobility disorders. This study evaluated the efficacy of treatment gallium maltolate (GaM) in rats infected with T. evansi in the acute and chronic phases of the disease and its influence on the enzyme and blood parameters. 48 animals (Rattus norvegicus) were divided into 8 groups (A-H) of 6 animals each, namely: A: (negative control) uninfected; B: acutely infected positive control; C: chronically infected positive control; D: acutely infected, treated with GaM for 7 days post infection (p.i.); E: acutely infected treated with GaM for 3 days before infection (b.i) and 7 days p.i.; F: chronically infected, treated with GaM for 7 days p.i.; G: chronically infected, treated with GaM for 3 days b.i. and 7 days p.i.; and H: uninfected treated with GaM for 10 days. Acute infected animals (B, D and E) had a progressive increase in parasitemia and were died or euthanized before completing treatment days (5th days p.i.) as they had high parasitemia (over 100 field trypanosomes in the blood smear). Thus, it can be concluded that GaM was not effective against an acute infection. In untreated chronically infected animals (C) the parasitemia also increased progressively and they were euthanized on the 7th day p.i.. The chronically infected and treated animals (F and G) showed low parasitemia and after treatment became negative, showing no trypanosomes in the bloodstream until the 50th day of the experiment. Thus, we conclude that GaM was effective against chronic infections. In uninfected and treated animals (H) hematological, biochemical and enzymatic parameters had no significant changes when compared to the negative control group (A) demonstrating the low toxicity of GaM., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Sarcocystis neurona , owing to its clinical importance in domestic animals, is currently one of the most studied agents, presenting a wide range of intermediate hosts that have not yet been described, mainly in wild fauna. Thus, the aim of this study was to describe the detection and molecular detection of S. neurona by amplification of the 18S rRNA region in the tissues of wild boars killed by boar control program in border Brazil Uruguay. A total of 79 samples of DNA from wild boar tissues from the LADOPAR/UFSM sampling bank were used, with Nested-PCR reactions being performed for amplification of the 18S rRNA region and the expected final product of 290 bp. Subsequently, the positive samples were subjected to restriction fragment length polymorphism (RFLP) technique with the restriction enzymes DdeI and HPAII . A second semi-Nested reaction was performed to obtain a larger sequence of nucleotides with amplification of the 18S region and the expected final product of 500 bp for S. neurona and Nested amplification ITS1 with product final of 367 pb. In 32 samples, it was possible to detect S. neurona both by nested Nested-PCR reaction and RFLP, and the presence of the agent was confirmed by sequencing, corresponding to 40.51% of the total tissues evaluated. This is the first report of the occurrence of this species of Sarcocystis in wild boars, and further studies evaluating the role of these animals as intermediate hosts, and in the epidemiology of this protozoan are necessary, as well as verifying the risk factors for infection., Competing Interests: Conflict of interestAuthors declare that there is no confict of interest., (© Indian Society for Parasitology 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Due to the proximity of humans to the countryside and the progressive increase in populations of invasive species, such as wild boars (Sus scrofa), the risk of disease spread is also exacerbated, some of which are zoonoses caused by protozoa. In the present study, 75 tissue/organ samples from 25 wild boars obtained from authorized hunting in the northern region of Rio Grande do Sul were evaluated to investigate the presence of Trypanosoma spp. using conventional PCR with specific primers and amplification of the ITS1 region for Leishmania spp. detection and species differentiation, multiplex PCR with kDNA minicircle amplification was performed. Trypanosoma spp. DNA was detected in 11 out of 25 hearts, representing 44% of the culled animals. Regarding the detection of Leishmania DNA, L. infantum was detected in one spleen sample, accounting for 4%, and L. amazonensis in one liver sample from the same animal, also representing 4% (1/25) of the samples. It is important to note that this wild boar, with detection for both L. amazonensis and L. infantum, also had Trypanosoma spp. DNA detected in a heart sample, indicating the potential of this species to have multiple infections with these agents. Furthermore, this is the first reported case of multiple infection in a wild boar with these agents. Therefore, the results obtained reinforce the risk posed by invasive species, especially wild boars, as potential sources of infectious agent dissemination and their role as possible reservoirs for numerous diseases., Competing Interests: Declaration of Competing Interest The authors declare no competing interests., (Copyright © 2023. Published by Elsevier B.V.)

Strangles is a contagious bacterial disease of horses caused by Streptococcus equi subspecies equi (SEE) that occurs globally. Rapid and accurate identification of infected horses is essential for controlling strangles. Because of limitations of existing PCR assays for SEE, we sought to identify novel primers and probes that enable simultaneous detection and differentiation of infection with SEE and S. equi subsp. zooepidemicus (SEZ). Comparative genomics of U.S. strains of SEE and SEZ (n = 50 each) identified SE00768 from SEE and comB from SEZ as target genes. Primers and probes for real-time PCR (rtPCR) were designed for these genes and then aligned in silico with the genomes of strains of SEE (n = 725) and SEZ (n = 343). Additionally, the sensitivity and specificity relative to microbiologic culture were compared between 85 samples submitted to an accredited veterinary medical diagnostic laboratory. The respective primer and probe sets aligned with 99.7 % (723/725) isolates of SEE and 97.1 % (333/343) of SEZ. Of 85 diagnostic samples, 20 of 21 (95.2 %) SEE and 22 of 23 SEZ (95.6 %) culture-positive samples were positive by rtPCR for SEE and SEZ, respectively. Both SEE (n = 2) and SEZ (n = 3) were identified by rtPCR among 32 culture-negative samples. Results were rtPCR-positive for both SEE and SEZ in 21 of 44 (47.7 %) samples that were culture-positive for SEE or SEZ. The primers and probe sets reported here reliably detect SEE and SEZ from Europe and the U.S., and permit detection of concurrent infection with both subspecies., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The aim of this study was to determine the presence of deoxyribonucleic acid (DNA) from Toxoplasma gondii, Sarcocystis spp. and Neospora caninum, in tissues of wild boars slaughtered in southern Brazil. A total of 156 samples were collected from different organs of 25 wild boars, and DNA from at least one of the protozoa investigated was detected in 79 samples. To differentiate between infectious agents, restriction fragment length polymorphism was performed using the restriction enzymes DdeI and HpaII. For N. caninum, conventional PCR was performed with specific primers. The DNA of at least one of the studied pathogens was detected in each animal: 26.58% for T. gondii, 68.36% for Sarcocystis spp. and 5.06% for N. caninum. Coinfection between T. gondii and Sarcocystis spp. occurred in 14 animals, between T. gondii and N. caninum in only one male animal, between Sarcocystis spp. and N. caninum in a female, while co-infection with the three agents was equally observed in only one male animal. Considering the high frequency of detection and its zoonotic risk, especially T. gondii, it appears that wild boars can be potential sources of transmission of infectious agents and the adoption of monitoring measures in these populations should be prioritized.

Lucilia cuprina is a vector of important diseases in humans and animals that causes myiasis in sheep, leading to enormous damage to the sheep sector. Chemical products are used to control these flies; however, there are reports of resistance in addition to these products causing toxicity to the environment, humans, and animals, so alternative controls have been studied to reduce these impacts. Pleurotus spp. are basidiomycete fungi and present bioactive compounds with medicinal properties. Due to the potential use of fungi to control Diptera, this study aimed to verify the activity of Pleurotus florida, P. ostreatus, and P. djamor in the control of larvae and adults of L. cuprina, as well as the effects of aqueous extracts of the fungi P. ostreatus, P. djamor, and P. florida on larvae and adults of L. cuprina. The aqueous extract from P. florida was the only one that showed larvicidal activity against L. cuprina, with a half-maximal effective concentration of 11.42 mg/mL. In the test with adult stages, 30 insects were used for each solution concentration, sprinkled with 1 mL of the solution. All aqueous extracts showed adulticidal activity at all concentrations, P. ostreatus showing the best results, with adult mortality ranging from 75.86 to 100%. Our results demonstrated an important larvicidal effect of P. florida and an adulticidal effect of all AE, with emphasis on P. ostreatus.

Prototheca bovis has been associated with several cases of mastitis in cattle but no record of intramammary infections has been reported in goats. This infection does not respond to available treatments and the disposal recommendation of affected animals cause great damage to the dairy industry. Alternatives for dealing with infections caused by Prototheca spp. are required worldwide. In vitro results suggest polypyrrole as promising molecule for combating this alga, because an algaecide effect was observed on tested Prototheca spp. isolates. Thus, this study evaluated goats as an experimental model for intramammary infection by P. bovis and a protocol for treating these animals with an intramammary polypyrrole solution. The possibility of P. bovis promoting an intramammary infection in goats was experimentally proven, demonstrating this species as an important model for studies involving algae mastitis. Furthermore, polypyrrole reduced the counts of Prototheca sp. in the analyzed samples, showing potential to fight this microorganism also in vivo. The results obtained in this study demonstrate the ability of P. bovis to colonize breast tissue in lactating goats and the highly soluble molecule of polypyrrole has potential use for the treatment of protothecosis., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Haemonchus contortus is a highly pathogenic and prevalent helminth that causes many deaths in sheep herds. Anthelmintics are usually employed to overcome this issue; however, they do not guarantee immediate and lasting efficacy because of the occurrence of drug-resistant parasites. Among substances that are used in scientific studies for parasitic control, essential oils are known to have different pharmacological properties. However, they demonstrate instability owing to several factors, and therefore, nanoemulsification is considered an alternative to control the instability and degradability of these compounds. The objective of this study was to evaluate the cytotoxicity of nanoemulsions containing essential oil of Eucalyptus globulus against the blood of healthy sheep and to verify their activity against the parasite H. contortus in sheep. The results presented adequate nanotechnological characteristics (diameter 72 nm, PDI 0.2, zeta -11 mV, and acidic pH) and adequate morphology. Further, the corona effect and cytotoxic profiles of the free oil and nanoemulsion against blood cells from healthy sheep were evaluated. The tests results did not present a toxicity profile. For evaluating efficacy, we observed an important anthelmintic action of the nanoemulsion containing oil in comparison to the free oil; the results demonstrate a potential role of the nanoemulsion in the inhibition of egg hatchability and the development of larvae L1 to L3 (infective stage). Based on these results, we developed an important and potential anthelmintic alternative for the control of the parasite H. contortus., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: Distiller's dried grains with solubles (DDGS) are coproducts of the biofuel industries that use corn as raw material. This cereal is commonly contaminated by mycotoxins, including fumonisins (FBs), which can pose a serious health threat to humans and animals. Corn DDGS are typically used as a protein-rich animal feed. As mycotoxins from the original cereal grains become concentrated in DDGS, mycotoxicological monitoring is highly required before their use as ingredient in the industry., Results: This work aimed to develop a methodology for predicting fumonisins B 1 (FB 1 ) and B 2 (FB 2 ) in corn DDGS using near-infrared reflectance spectroscopy (NIRS) technology associated with chemometric methods. One hundred and ninety corn DDGS samples originating from Brazilian ethanol plants and feed mills were included in this assessment. Two datasets were created: one for calibration (132 samples) and another for external validation (58 samples). Partial least squares regression and a cross-validation approach were applied to build the models. Liquid chromatography coupled to tandem mass spectrometry was used as the reference methodology. Calibration results of correlation coefficient and residual prediction deviation for FB 1 and FB 2 were, respectively, 0.90 and 0.88; and 2.16 and 2.06., Conclusion: Values of the external validation dataset were compared and no statistical difference was found between groups, indicating a satisfactory predictive ability and confirming the potential of NIRS to predict fumonisins in corn DDGS. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

Microbiological diagnosis of equine respiratory infections is essential for disease management. However, reliable diagnosis can be a challenge due to colonization of the upper respiratory tract (URT) by a diverse microbial population, and because there is a lack of studies with samples from healthy animals. Aiming to guide adequate URT culture, this work reports culturable microbial population from the URT of 1,010 apparently healthy horses from 341 farms in Southern Brazil and identifies the putative presence of pathogenic microorganisms. Nasal swabs were cultured in 5% blood agar, and the isolates were phenotypically identified to genus level. A diverse respiratory microbial population was found, mostly composed of Gram-positive bacteria, including Staphylococcus spp., Bacillus spp., Streptococcus spp. and Corynebacterium spp. The microbiological profile from the nasal cavity of 911 horses was described, with the five most common profiles being: (1) Staphylococcus sp. + Gram-negative bacilli (12.67%), (2) Staphylococcus sp. in pure culture (12.47%), (3) Staphylococcus sp. + Bacillus sp. (10.10%), (4) Gram-negative bacilli in pure culture (6.93%), and (5) Staphylococcus sp. + Bacillus sp. + Gram-negative bacilli (6.73%). Streptococcus equi equi and Rhodococcus equi were detected in 34 horses (3.37%), demonstrating the presence of pathogenic bacteria along with commensal microorganisms in apparently healthy animals. The disclosed data may guide and facilitate microbiological diagnosis of URT infection in horses. The significant presence of Gram-negative bacilli was evidenced, as well as the occurrence of relevant pathogens, such as S. equi equi and R. equi, thus helping to improve diagnosis and antimicrobial therapy., (Copyright © 2022. Published by Elsevier Inc.)

The etiology of otitis in dogs and cats is multifactorial and complex, involving bacterial and fungal pathogens. As empiric antimicrobial prescription is a common practice when treating such cases, antimicrobial resistance may represent a complicating factor. The aim of this study was to describe microbiological features and susceptibility profiles of pathogens associated with 142 cases of external otitis, comprising 138 dogs and 4 cats.. The specimens were processed to identify bacterial and fungal etiologies following standard microbiological methods. Antimicrobial susceptibility was determined in vitro against 15 antibiotics and 3 antifungals. Further, Staphylococcus spp. isolates were screened for the detection of β-lactamase enzymes using cefinase paper discs. Pseudomonas spp. and isolates from Enterobacteriaceae family were screened for colistin (Polymyxin E) resistance and for the mcr-1-mediated colistin resistance gene by PCR. The presence of mixed cultures of Enterobacteriaceae, Pseudomonas spp. and Staphylococcus spp., and co-infections with Malassezia spp., emphasizes the polymicrobial etiology of external otitis in small animals. Emerging rates of multidrug resistance observed in almost 50% of the isolates may alert for a near future of challenging veterinary cases unresponsive to first-line antimicrobials. In addition, these results highlight a potential public health concern of multidrug resistant bacteria, given the proximity of pets and their owners. This study addressed central aspects of external otitis, providing microbiologists and clinicians updated information on the etiology and treatment of challenging cases of multidrug resistant bacteria. It also provides priceless surveillance value in monitoring resistant bacteria in small animals., (Copyright © 2022. Published by Elsevier Ltd.)

Cyathostomins are common intestinal nematodes that parasitize horses and can affect animal health and welfare. Alternative treatments are essential for sustainable parasite control, reducing the indiscriminate use of anthelmintics and their impacts on the environment, animals, and humans. Pleurotus spp. are edible medicinal mushrooms that possess several therapeutic properties, including antiparasitic and nematocidal activities. This study aimed to evaluate the antioxidant and nematocidal activity of aqueous extracts of three oyster species (P. florida, P. ostreatus, and P. djamor) on cyathostomine eggs in vitro. P. florida showed the highest activity against test organisms, inhibiting the hatching of eggs in up to 92.19% at the highest concentration (10%) of aqueous extract, with a mean lethal concentration (LC50) of 2.13%. In the aqueous extracts of P. ostreatus and P. djamor, nematocidal activity was reduced, inhibiting egg hatching by 55.46% (LC50, 7.44%) and 23.67%, respectively, at a concentration of 10%. P. florida showed higher antioxidant activity than that of the other species tested. Thus, P. florida may be considered a potential alternative for biological control in in vivo experiments with cyathostomins.

The aim of the current study is to investigate whether silver catfish (Rhamdia quelen) individuals exposed to commercial formulation of the chlorantraniliprole insecticide used in rice crops present changes in biochemical parameters. Fifty-four (54) silver catfish individuals were distributed in six units per tank (n = 6/repetition; triplicate/treatment) and subjected to the following treatments: T1-control, without insecticide; T2 (0.02 µg/L of insecticide) and T3 (0.20 µg/L of insecticide). Exposure time lasted 24 or 96 h, and it was followed by 96 h recovery in pesticide-free water. Results have indicated biochemical changes in cortisol, glucose, lactate and plasma protein levels, as well as few ionic changes in animals' gills during the exposure and recovery periods. Chlorantraniliprole incidence in water resulted in some biochemical changes in silver catfish specimens' plasma and gills throughout the acute exposure protocol (sub-lethal dose). Thus, chlorantraniliprole insecticide has caused osmoregulatory and/or biochemical imbalance in the investigated species under the herein adopted laboratory conditions; these changes did not get back to normal levels even after specimens were left to recover for 96 h in clean water., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

This study aimed to evaluate the applicability and efficiency of Near-Infrared Spectroscopy (NIR) by using dispersive NIR and Fourier Transform NIR to analyse 267 samples of Brazilian wheat flour contaminated with deoxynivalenol (DON). For this, Partial Least-squares Discriminant Analysis (PLS-DA) and Principal Component Analysis-Linear Discriminant Analysis (PC-LDA) were used as discriminatory methods. Next, the samples were classified according to the maximum tolerated limits (MTL) for DON in Brazil, 750 μg kg -1 , and two groups were established for the calibration set: category A (≤450 μg kg -1 ), non-contaminated or below the MTL; and category B (>450 μg kg -1 ), contaminated or above the MTL. Validation samples through PLS-DA showed correct classification rates in the range of 85-87.5% and presented a 10-15% error; for PC-LDA, the hit rate was over 85% with an error of 10-15%. The present findings demonstrate that NIR is an excellent alternative method to classify wheat flour samples according to DON content.

The impact of deoxynivalenol (DON) upon intestinal tissue of broilers was assessed by using jejunal explants in Ussing chambers and analyzing histopathological and immunohistochemical parameters; this system was also applied to evaluate the efficacy of an antimycotoxins additive (AMA). The explants were subjected to the following treatments within each experiment for 120 min: Experiment 1) T1 (control) - buffer solution, and T2 - 10 mg/L DON; and Experiment 2) T1 (control) - buffer solution, T2 - 10 mg/L DON, T3 - AMA (0.5%), and T4 - 10 mg/L DON + 0.5% AMA. In Experiment 1, DON triggered a reduction in the size of enterocytes as well as of their nuclei, an increase in cytoplasmic vacuolization and apical denudation of villi. Apoptotic cells count was also greater in DON-exposed explants. In Experiment 2, the AMA mitigated DON harmful effects; cytoplasmic vacuolization of enterocytes was reduced and the size of their nuclei was preserved. The additive also promoted a partial decrease in microvillus integrity, in size of enterocytes and in apoptotic cells count. The tested ex vivo model demonstrated the impact of DON upon the intestine as well as the efficacy of the AMA against its damaging effects., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Fumonisins (FBs) and zearalenone (ZEN) are mycotoxins which occur naturally in grains and cereals, especially maize, causing negative effects on animals and humans. Along with the need for constant monitoring, there is a growing demand for rapid, non-destructive methods. Among these, Near Infrared Spectroscopy (NIR) has made great headway for being an easy-to-use technology. NIR was applied in the present research to quantify the contamination level of total FBs, i.e., fumonisin B1+fumonisin B2 (FB1+FB2), and ZEN in Brazilian maize. From a total of six hundred and seventy-six samples, 236 were analyzed for FBs and 440 for ZEN. Three regression models were defined: one with 18 principal components (PCs) for FB1, one with 10 PCs for FB2, and one with 7 PCs for ZEN. Partial least square regression algorithm with full cross-validation was applied as internal validation. External validation was performed with 200 unknown samples (100 for FBs and 100 for ZEN). Correlation coefficient (R), determination coefficient (R2), root mean square error of prediction (RMSEP), standard error of prediction (SEP) and residual prediction deviation (RPD) for FBs and ZEN were, respectively: 0.809 and 0.991; 0.899 and 0.984; 659 and 69.4; 682 and 69.8; and 3.33 and 2.71. No significant difference was observed between predicted values using NIR and reference values obtained by Liquid Chromatography Coupled to Tandem Mass Spectrometry (LC-MS/MS), thus indicating the suitability of NIR to rapidly analyze a large numbers of maize samples for FBs and ZEN contamination. The external validation confirmed a fair potential of the model in predicting FB1+FB2 and ZEN concentration. This is the first study providing scientific knowledge on the determination of FBs and ZEN in Brazilian maize samples using NIR, which is confirmed as a reliable alternative methodology for the analysis of such toxins., Competing Interests: The authors have read the journal’s policy, and the authors of this manuscript have the following competing interests to declare: AOM is a paid employee of Pegasus Science. This does not alter our adherence to PLOS ONE policies on sharing data and materials. There are no patents, products in development or marketed products associated with this research to declare.

Antimicrobial-resistant bacteria have been frequently isolated from canine otitis. Photodynamic therapy using porphyrins as photosensitizing molecules is an alternative therapy against microorganisms in localized infections. Therefore, we evaluated the antibacterial activity of two tetra-cationic porphyrins (H 2 TMeP and ZnTMeP) against gram-positive and -negative bacteria isolated from canine otitis, as well as its putative action mechanism. For this, two gram-positive and two gram-negative bacteria frequently detected in cases of canine otitis (coagulase-positive and -negative staphylococci [CPS and CNS], Pseudomonas aeruginosa, and Proteus mirabilis) were used in antibacterial activity assays. Each porphyrin at a non-cytotoxic dose was incubated with a fixed concentration of each bacterium and exposed to white-light irradiation for 0, 30, 60, and 90 min. Clinical isolates of CPS and CNS were photo-inactivated after 30 min of white-light exposure by both porphyrins (p < 0.05). Gram-negative bacteria were also photo-inactivated after 30 min (p < 0.05), reaching complete inactivation after 60 and 90 min of white-light irradiation by H 2 TMeP and ZnTMeP, respectively. Antibacterial assays using standard bacterial strains (ATCCs) demonstrated similar results to those obtained with clinical isolates, except for P. aeruginosa, which was completely inactivated by ZnTMeP at 60 min, and the absence of a significant reduction in P. mirabilis concentration when irradiated for 30 min. Similar assays were conducted using reactive oxygen species scavengers showing that the putative mechanism for bacterial inactivation is through the production of singlet oxygen species. These results indicated that H 2 TMeP and ZnTMeP tetra-cationic porphyrins were effective against bacteria isolated from canine otitis., (Copyright © 2020 Elsevier B.V. All rights reserved.)

It is well documented today that pesticides, used in crop production, may modulate the immune system of healthy fish. However, there is still only limited information regarding the effects of these anthropogenic stressors in conjunction with natural stressors (pathogens), on the innate immune responses of freshwater fish. Thus, the aim of this investigation was to compare the innate immune response of two groups of fish (Rhamdia quelen), naturally infected with Aeromonas hydrophila, exposed and unexposed to a non-lethal concentration of chlorantraniliprole (CAP) insecticide (0.0 and 1.3 μg/L/24 h). Unhealthy fish exposed to CAP showed significant higher total leukocyte counts and neutrophils percentage compared to non-exposed infected fish). However, the monocytes and eosinophils percentage significantly decreased in fish exposed to CAP. Furthermore, lysozyme activity values measured in plasma, skin mucus, gill and intestine significantly reduced in fish exposed to CAP. The CAP-induced immunomodulation may interfere on the ability of the animal to heal or fight the infection, and possible contribute to the spread of bacterial infection in fish production or environment., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A mycotoxicological survey was conducted in breakfast (n = 172) and infant (n = 43) cereals commercialized in Brazil. Samples were collected in 2018 for analyses of: aflatoxins (AFs) B 1 (AFB 1 ), B 2 , G 1 and G 2 ; fumonisins (FBs) B 1 (FB 1 ) and B 2 ; zearalenone (ZEN); the trichothecenes (TRCs) deoxynivalenol (DON), T-2 toxin, HT-2 toxin, nivalenol, fusarenon X, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol and diacetoxyscirpenol; and ochratoxin A. FB 1 was the most prevalent metabolite in breakfast cereals, being detected in 26.7% of the samples (mean 105 μg/kg); ZEN had the second highest positivity, 14.8% (mean 17 μg/kg), followed by DON with 10% (mean 44 μg/kg). In infant cereals, FB 1 also had the highest incidence, 27.8% (mean 55 μg/kg), followed by DON with 10.3% (mean 36 μg/kg) and ZEN with 6.9% (mean 3 μg/kg). Mycotoxins contamination was found in 31.4% (n = 54) of the breakfast cereals and in 18.6% (n = 8) of the infant cereals. In these positive samples, co-occurrence of two or three mycotoxins was detected in 31.5% (n = 17) of the breakfast cereals and in 25% (n = 2) of the infant cereals. The mycotoxins found co-contaminating the breakfast cereals belong to the genera Aspergillus and Fusarium; ZEN, followed by AFB 1 , were the most prevalent ones. As for the infant cereals, the associated fungal metabolites are produced by the genus Fusarium; the highest incidence was seen for ZEN. Low contamination and positivity of mycotoxins were found herein; nonetheless, in some samples these substances were present at levels which transgress those preconized in the Brazilian legislation. Therefore, mycotoxicological monitoring of this type of product throughout the nation is crucial in order to identify the potential risk to which the Brazilian population is exposed, particularly the children., Competing Interests: Declaration of competing interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Data about phylogenetic classification of Escherichia coli colonizing calves, lambs and foals are routinely neglected and restricted to outdated methodologies, even in the context of antimicrobial susceptibility (AS) testing. Thus, the aim of this study was to determine the phylogenetic diversity and the AS profile of E. coli colony-forming units (CFUs) from faecal samples of healthy animals. Five CFUs of E. coli were randomly selected from each faecal culture of calves (n = 13), foals (n = 13) and lambs (n = 13), totalizing 195 CFUs phylo-typed by quadruplex PCR. The AS profile of five CFUs from 15 samples (five from each animal species; n = 75 isolates) against nine drugs was determined by agar diffusion test. We found E. coli belonging to all phylo-groups already described, except D group, with the predominance of B1 (65% CFUs; 126/195) in the three-animal species sampled. Most faecal samples of calves (77%; 10/13) and foals (69%; 9/13) harboured both pathogenic and nonpathogenic E. coli. All faecal samples showed CFUs with diverse AS profile, highlighting the ineffectiveness of tetracycline, sulphonamide and ampicillin. As a key point, our data reinforce the importance to select at least four E. coli CFUs for AS testing. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides relevant data about the high phylogenetic and antimicrobial susceptibility diversity observed in Escherichia coli colony-forming units (CFUs) from a bacteriological culture of faeces from healthy calves, foals and lambs. The selection pressure exerted by the herd treatment may directly impact the intestinal microflora of animals that have never been treated. Finally, we emphasize the importance of Clinical Laboratory Standards Institute guidelines and we recommended to analyse at least four E. coli CFUs to determine, in particular, the antimicrobial susceptibility profile of faecal isolates, independent of the animal's health status., (© 2019 The Society for Applied Microbiology.)

The ability of Rhodococcus equi to survive in macrophages and cause pneumonia in foals depends on vapA and rhbC genes, which produce the virulence-associated protein A (VapA) and the rhequichelin siderophore, respectively. Virulent R. equi acquires Fe from transferrin by unknown mechanisms. Our objectives were to determine the role of GAPDH in Fe homeostasis, to further characterize GAPDH, rhbC, and vapA expression under iron homeostasis, and to document the occurrence of rhbC gene in R. equi isolates. Therefore, vapA + R. equi was cultured under excessive, physiologic, and restricted iron concentrations, and quantitative culture and gene expression were performed. The relative expression of GAPDH, rhbC, and vapA after 48 h of culture were analyzed by qPCR. To determine the rhbC occurrence, total DNA was extracted from R. equi isolated from foals with clinical rhodococcosis (n = 22), healthy horses (feces, n = 16; nasal swab, n = 9), soil (n = 6), and 2 ATCC reference strains. Conventional PCR was performed to identify genus/species, vapA, and rhbC genes. Iron restriction proportionally decreased R. equi growth rates, and induced high expression of both GAPDH and vapA. The putative role of GAPDH in R. equi iron homeostasis should be further investigated. rhbC was significantly up-regulated under both Fe excess and critical starvation. The rhbC gene was identified in all clinical isolates and soil, but it was absent in 2 isolates from healthy horses, suggesting that rhequichelin is not required for R. equi nasal and intestinal colonization., Competing Interests: Declaration of competing interest None., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

This study assessed the hematological, enzymatic and osmoregulatory responses of silver catfish ( Rhamdia quelen ) exposed to sublethal concentrations (1.125 and 3.750 µg/L) of a commercial thiamethoxam-containing insecticide used on rice crops. Groups of 6 fish per tank (in triplicate, n = 3, total 54 fish) were exposed for up to 96 h to different concentrations of the compound. After this period, fish were placed in clean water for 48 h. Two fish from each tank (6 per treatment) that had been exposed to the insecticide for 24 h were anesthetized with eugenol and blood was collected to evaluate hematological and biochemical parameters. Blood, liver and muscle were collected for determination of metabolic parameters, plasma cortisol, Cl - , Na + and K + levels and H + -ATPase and Na + /K + -ATPase activity in the gill. H + -ATPase activity was higher in fish exposed to 1.125 µg/L insecticide at 24 h compared to control (0.0 µg/L). Differences in cortisol levels were evidenced throughout the experimental period. These results indicated that exposure to the insecticide changed the hematological, biochemical and metabolic profile of the animals, suggesting concern about environmental safety. Therefore, we discourage the use of this pesticide in areas that come into contact with water bodies inhabited by fish.

Many important studies on resistance reversion, anthelmintic efficacy and, especially, new molecules with antiparasitic effects are performed in laboratories using gerbils (Meriones unguiculatus) as the experimental model. This study aimed to evaluate the use of corticosteroids (dexamethasone and methylprednisolone acetate) in gerbils experimentally infected with different doses of infective larvae (sheathed or exsheathed) of Haemonchus contortus. In the first experiment, 28 gerbils were divided into seven groups infected by 2-6 × 103 larvae, with or without immunosuppression using corticosteroids. In the second experiment, eight gerbils were divided into two groups infected by 2 × 103 sheathed or exsheathed larvae. For the third assay, seven immunosuppressed gerbils were infected with 2 × 103 sheathed larvae and were killed 15 days post infection (PI). The highest number of parasites was recovered from methylprednisolone-immunosuppressed animals. We observed red and white blood cell alterations and biochemical parameters in infected animals that had undergone immunosuppression with methylprednisolone. We highlight that in the first and second experiments a satisfactory number of worms was recovered using sheathed larvae and immunocompetent animals. When exsheathed larvae were used, the number of worms recovered was unsatisfactory. A considerable larval burden was recovered from immunosuppressed gerbils 15 days PI, and body weight did not influence establishment of larvae.

The aim of this study was to evaluate in vitro the efficacy of cordycepin and pentostatin (alone or combined) against Trypanosoma cruzi, as well as the therapeutic efficiency of protocols of cordycepin and pentostatin combinations in mice experimentally infected with T. cruzi. In vitro, the cordycepin (3'-deoxyadenosine) and pentostatin (deoxycoformycin) exerted potent trypanocidal effect against T. cruzi (Colombian strain), similarly to benznidazole, which is the reference drug. For epimastigotes, the lethal dose of cordycepin capable of killing 50% (LD 50 ) and 20% (LD 20 ) of the parasites was 0.072 and 0.031 mg/mL, respectively and for trypomastigotes was 0.047 and 0.015 mg/mL, respectively. The combined use of cordycepin and pentostatin resulted in a LD 50 and LD 20 for epimastigotes of 0.068 and 0.027 mg/mL, respectively, as well as 0.056 and 0.018 mg/mL for trypomastigotes, respectively. In vivo, the combined use of cordycepin and pentostatin did not show the expected curative effect, however it was able to control the parasitema in the peak period. In summary, the combination of cordycepin and pentostatin showed no curative effect in mice infected by T. cruzi, despite the in vitro reduction of epimastigotes and trypomastigotes., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Purpose: To address more information about changes in commensal Escherichia coli during virus intestinal infection, we characterized 30 faecal E. coli isolates from calves (21 to 60 days old) with diarrhea due to rotavirus and coronavirus, which received, before diagnosis, tetracycline, gentamicin and enrofloxacin drugs., Methodology: Clermont's phylogenetic classification; presence of genes for curli, cellulose, fimbriae (F4, F5, F6, F18, F41); and antimicrobial susceptibility were used to characterize the isolates. Disk diffusion technique and PCR were used as methodologies., Results: E. coli isolates from calves with diarrhea were phylogenetically classified as B1 (70%, 21/30), B2 (3.33%, 1/30), C (3.33%, 1/30), D (3.33%, 1/30), E (13.33%, 4/30) and unknown (6.7 %; 2/30), whereas E. coli isolates from the control group were classified only as B1 (83.3%, 25/30), E (10 %; 3/30) and unknown (6,7 %; 2/30). E. coli isolates from calves with diarrhea showed a much higher resistance profile with 16 (53.3%) multiresistant isolates. Only isolates (30%-9/30) from diarrheic calves were also positive for fimbriae, specifically 16.7% (5/30) for F5 and 13.3% (4/30) for F18., Conclusion: To sum up, E. coli isolates from calves with diarrhea showed differences in relation to the control group, confirming changes in commensal E. coli during virus intestinal infection. It can be emphasized that some care should be taken to manage diarrheic calves: the pathological agent must be diagnosed prior to treatment; antibacterial treatment should be with antimicrobials with a different mechanism of action; and finally, treated animals should be maintained separately from others because they can carry micro-organisms with a resistant profile.

Background: Rhodococcus equi is an important cause of foal pneumonia. While its isolation from different sources has been widely evaluated, there is a need to better understand the R. equi epidemiology from samples of the nasal cavity of healthy horses., Objectives: To determine the prevalence of R. equi from the nasal cavity of healthy horses, along with its virulence profile, antimicrobial susceptibility and environmental variables associated., Study Design: Cross-sectional study., Methods: Swabs from the nasal cavity of 1010 apparently healthy horses from 341 farms were submitted for bacteriological analyses. The identity and virulence profile of the R. equi isolates were assessed by multiplex PCR; antimicrobial susceptibility was determined by the disk-diffusion method. The occurrence of R. equi was calculated at the level of both animal and farm. The association of seven specific environmental factors with R. equi isolation was assessed using logistic regression and by a spatial scan statistical method to determine the presence of local clusters., Results: Antimicrobial-sensitive R. equi was isolated from 10 (1%) of 1010 horses ranging between 3 and 29 years old. Ten farms (3%) had at least one positive horse. Only one R. equi isolate (10%) was classified as virulent. Red-Yellow Argisol (PVA/PV) soils were significantly associated with R. equi isolation (odds ratio (OR) 8.02; CI 95% , 1.98-32.50, P = 0.01), and areas with well-drained soil were less likely to be test positive (OR 0.85; CI 95% , 0.76-0.96, P = 0.03)., Main Limitations: The use of culture-based method instead of PCR-based assay and the lack of soil sampling., Conclusions: Antimicrobial-sensitive R. equi may be considered a minor part of the normal bacterial flora in the nasal cavity of healthy and immunologically functional horses breeding on pasture. Further studies are warranted to determine if soils rich in iron and well-drained are, in fact, associated with the occurrence of R. equi., (© 2018 EVJ Ltd.)

Aims: This study investigated the antibacterial activity of five phytochemicals (carvacrol, citral, eugenol, linalool and thymol) alone or in combination with florfenicol or oxytetracycline against bacteria isolated from silver catfish (Rhamdia quelen). We also analysed the potential of these compounds to inhibit biofilm formation and haemolysis caused by the bacteria., Methods and Results: Bacteria were tested with antimicrobials to calculate the multiple antibiotic resistances. The checkerboard assay was used to evaluate a putative synergy between five phytochemicals and antimicrobials against the strains isolated. The biofilm formation inhibition assay was performed with phytochemicals and antimicrobials, and the haemolysis inhibition assay was performed with the phytochemicals. Carvacrol, eugenol and thymol were the most effective phytochemicals. Three combinations (linalool with florfenicol or oxytetracycline against Aeromonas hydrophila and citral with oxytetracycline against Citrobacter freundii) demonstrated synergy in the checkerboard assay. All phytochemicals inhibited biofilm formation and haemolysis activity., Conclusion: The tested phytochemicals showed satisfactory activity against fish pathogenic bacteria., Significance and Impact of the Study: The phytochemicals did not present antagonistic interactions with the antimicrobials, allowing their combined use, which may contribute to a decrease in the use of conventional drugs and their residues in aquatic environment., (© 2018 The Society for Applied Microbiology.)

The effects of pre-transport handling and addition of essential oil of Myrcia sylvatica (EOMS) during transport on stress pathways activation in Rhamdia quelen were investigated. Fish (n=400, 25.2±2.9g) were captured in production ponds and transferred to 100-L tank (density 100g L -1 ). After 24h, 10 fish were sampled (before transport group). The remaining fish were placed in plastic bags (n=30 or 32 fish per bag, density 150g L -1 ) containing 5L of water (control), ethanol (315μLL -1 , vehicle) or EOMS (25 or 35μLL -1 ), in triplicate, transported for 6h and sampled (n=10 animals per group). Indicators of stress and metabolism, as well as mRNA expression of brain hormones were evaluated. Previously, full-length cDNAs, encoding specific corticotropin-releasing hormone (crh) and proopiomelanocortins (pomca and pomcb), were cloned from whole brain of R. quelen. Crh expression increased after 24h of capture and handling, whereas cortisol and glucose plasmatics enhanced their values in the control group. Transport with EOMS reduced plasma cortisol and lactate levels, while ethanol and EOMS groups increased Na + /K + -ATPase gill activity compared to control. Gene expression of crh, pomcb, prolactin and somatolactin mRNAs were lower after transport with EOMS compared to control. EOMS was able to mitigate the stress pathways activation caused by transport, maintaining a balance in body homeostasis. Thus, EOMS is recommended as sedative in procedures as transport and the pre-transport handling requires greater attention and use of tranquilizers., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Rhodococcus equi is an opportunistic, intracellular pathogen that causes pyogranulomatous pneumonia in foals and immunocompromised people. Currently, there is no experimental model of R. equi pneumonia other than intra-bronchial experimental infection of foals with R. equi, which is labor-intensive and costly. This study's objective was to develop a guinea pig (GP) model of R. equi pneumonia that would facilitate development of novel approaches for controlling and preventing this disease. Guinea pigs were infected with either 10 1 , 10 2 , 10 3 , or 10 4 colony forming units (CFUs) of a virulent strain of R. equi using a Madison aerosol chamber, or 10 6 or 10 7 CFUs of this strain intratracheally. Animals were monitored daily for clinical signs of pneumonia, and were euthanized and necropsied on days 1, 3, 7, or 35 post-infection (PI). Lung homogenates were plated onto selective agar to determine bacterial load. No clinical signs of disease were observed regardless of the inoculum dose or infection method. No bacteria were recovered from GPs euthanized at 35 days PI. Histology and immunostaining of T-cells, B-cells, and macrophages in lungs showed that inflammatory responses in infected GPs were similarly unremarkable irrespective of dose or route of infection. Guinea pigs appear to be resistant to pulmonary infection with virulent R. equi even at doses that reliably produce clinical pneumonia in foals., (Copyright © 2017 Elsevier B.V. All rights reserved.)

This study evaluated the activity of leaf essential oil (EO) from Hyptis mutabilis as well as its major constituent, (-)-globulol, in infections by the parasite Ichthyophthirius multifiliis (ich). Effects on hematological, biochemical, and immunological parameters of silver catfish, Rhamdia quelen, exposed to the same samples also were evaluated. In the first experiment, naturally infected fish were treated with EO (0, 10, and 20 mg · L -1 ) and ethanol, using several methods of exposure. Fish mortality and the number of trophonts per fish were assessed after 48 and 96 hr. Hour-long daily baths resulted in optimal survival, so this methodology was used for the second experiment, in which infected animals were exposed to (-)-globulol at 2.5 and 5 mg · L -1 . The most effective concentrations in Experiments 1 and 2 were chosen for Experiment 3, in which healthy animals were subjected to hour-long daily baths with EO (20 mg · L -1 ) or (-)-globulol (2.5 mg · L -1 ). Additionally, an in vitro experiment was performed with EO and globulol at the same concentrations of the in vivo test. EO and (-)-globulol increased the survival of fish infected with ich and altered certain hematological and biochemical parameters. After 4 days, levels of hematocrit, erythrocytes, and leukocytes increased significantly in healthy animals exposed to EO. Exposure to (-)-globulol increased leukocyte number alone. No significant differences in nonspecific immunological parameters were detected when treated groups were compared to controls, but the leukocytosis observed in EO- and globulol-treated healthy animals indicates that EO and (-)-globulol increased innate immunity in these fish. An in vitro antiparasitic effect was observed in both samples.

The aim of this study was to evaluate the efficacy of 3'-deoxyadenosine and deoxycoformycin combination in the treatment of mice infected by T. cruzi, as well as to verify the influence of the treatment on purinergic enzymes. Heart and serum samples were collected from 60 mice (30 infected and 30 uninfected) at day 12 post-infection. To verify treatment efficacy, parasitemia was monitored, and the treatment with 3'-deoxy adenosine and deoxycoformycin combination was able to reduce it, but had no curative effect on mice. Seric activities of NTPDase (ATP and ADP substrate) and ADA were increased significantly in untreated mice infected by T. cruzi compared to the negative control, as well as mice treated with 3'-deoxyadenosine and deoxycoformycin (alone or combined) modulated the activity of NTPDase (ATP and ADP substrate), preventing them from increasing in infected animals (activity similar to healthy animals). Treatment with deoxycoformycin alone and associated with 3'-deoxyadenosine modulated the activity of ADA preventing them from increasing in infected animals. However, seric activities of ADA in mice treated with 3'-deoxyadenosine (cordycepin) alone does not modify the ADA activity compared with infected and non-treated mice. However, the 5'-nucleotidase activity decreased significantly in infected untreated animals and the same occurred in infected and treated animals with deoxycoformycin and 3'-deoxyadenosine. However, treatment with deoxycoformycin associated with 3'-deoxyadenosine preventing them from decreasing the 5'-nucleotidase activity. Therefore, we conclude that the treatments did not have curative success for mice infected by T. cruzi. However, the treatments were able to modulate the purinergic enzymes during the infection by T. cruzi, which may contribute to reduce the inflammatory damage in heart., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Moraxella bovis is historically known as the primary agent of infectious bovine keratoconjunctivitis (IBK). However, Moraxella bovoculi and Moraxella ovis are also reported to be involved in the pathogenesis of IBK, therefore, these three species should be included in the development of a new vaccine with a broad-spectrum protection against the disease natural challenge. In this study we investigated the antigenic properties of clinical isolates and reference strains of M. bovis, M. bovoculi and M. ovis using a novel in vitro approach for vaccine evaluation based on two techniques, flow cytometry and western blotting (WB). Here, we demonstrated that rabbit antisera produced against reference M. bovis strain and commercial bacterin showed low number of IgG with capacity to recognize a panel of heterologous strains composed by M. bovoculi and M. ovis. On the other hand, the antisera generated against two clinical isolates of M. ovis (Mov2 and Mov3) presented high cross-reactivity levels against all M. ovis and M. bovis strains evaluated. Similarly, the antisera against Mbv3 (clinical isolate of M. bovoculi) had high levels of IgG associated on the surface of all M. bovoculi strains and most of the M. ovis strains analyzed. The WB analysis demonstrated that Moraxella spp. has multiple immunogenic antigens and most of them are shared between the three species. Based on the cross-reactivity analysis and considering the relative number of IgGs associated on the bacterial surface, we suggest that a multivalent vaccine including Mbv3, Mov2 and Mov3 strains may provide a strong and broad protection against all strains involved in IBK outbreaks., (Copyright © 2017 Elsevier B.V. All rights reserved.)

In the present study, Litomosoides silvai parasitizing Akodon montensis in the southern region of Brazil is reported for the first time. New morphological information is provided for some structures of this nematode species, such as a flattened cephalic extremity, presence of two dorsal cephalic papillae, female tail with a constriction at its tip, "s" shaped vagina, spicules characteristic of the carinii species group and microfilaria tail constricted at the tip. This nematode was found parasitizing the thoracic cavity with a prevalence of 10% (2/20), mean intensity of 4 (6/2), mean abundance of 0.4 (8/20) and range of infection of 2-6 specimens per host, in southern Brazil. This occurrence of L. silvai in A. montensis is a new geographical record for southern Brazil, in the Upper Paraná Atlantic Forest ecoregion of the northwestern region of Rio Grande do Sul, which is part of the Atlantic Forest biome.

The effect of dissolved organic carbon (DOC) against the burden of low pH was assessed in silver catfish Rhamdia quelen through the gills. A commercial humic acid (HA) was used as the source of DOC at 0 (control), 10, 25, and 50 mg/l. For each HA concentration, two pH levels were tested: 6.5 (control) and 5.5. After 40 days of exposure, the gills were removed and morphological variables were analyzed through light and scanning electronic microscopy. The low water pH caused a reduction in the length of filaments, number of lamellae, and gill respiratory surface area (GRSA) and an increase in chloride cells (CCs) number in the filament epithelium. When HA was added to the test water, GRSA increased and CCs proliferated in lamellae, suggesting ionic and respiratory disturbances. Scanning electronic microscopy revealed that CC morphometric variables, that is, apical area, fractional area, and density, were undisturbed by the pH reduction, but increased in the presence of HA. This study indicates that the commercial HA did not protect silver catfish against low pH stress. Instead, it caused changes that may affect vital processes such as ion regulation and ventilation and, consequently, reduce growth., (© 2017 Wiley Periodicals, Inc.)

Coagulation disorders have been described in Chagas disease with thrombocytopenia as an important event. Several mechanisms may be related to this pathogenesis, such as enzymes of the purinergic system, purine, and receptors involved in the regulation and modulation of physiological events related to hemostasis. Therefore, the aim of this study was to evaluate the activities of E-NTPDase, E-5'nucleotidase, and ecto-adenosine deaminase (E-ADA) in platelets of mice experimentally infected by Trypanosoma cruzi. Twelve female mice were used, divided into two groups (n = 6): uninfected and infected. Mice of infected group were intraperitoneally inoculated with 10 4 trypomastigotes of T. cruzi (strain Y). On day 12 post-infection (PI), blood samples were collected for quantitation and separation of platelets. A significant reduction in the number of platelets of infected mice (P < 0.05) was observed. The activities of E-NTPDase (ATP and ADP substrates), E-5'nucleotidase, and E-ADA in platelets increased significantly (P < 0.05) in mice infected by T. cruzi compared with uninfected animals. A negative correlation (P < 0.01)was observed between the number of platelets and ATP hydrolysis (r = -0.64), and ADP hydrolysis (r = -0.69) by E-NTPDase. Therefore, there is a response from the purinergic system activating ecto-enzymes in platelets of mice T. cruzi infected, as a compensatory effect of thrombocytopenia.

Pythium insidiosum is an aquatic oomycete that causes pythiosis, an important and severe disease of difficult treatment that affects humans, domestic and wild animals. This infection is often described in horses in Brazil and humans in Thailand. In clinical practice, we have observed many cases that do not respond to available therapies, indicating the need to explore alternative therapeutic approaches. In this sense, studies using metal compounds in conjunction with available antimicrobial agents have been demonstrated greater antimicrobial activity. Thus, in this research, we tested in vitro activities of metallic compounds containing cadmium, lead, copper, manganese, or zinc against 23 isolates of P. insidiosum. The assays were performed by broth microdilution based on CLSI M38-A2 document. The minimum inhibitory and fungicidal concentrations were established for all isolates. Copper acetate and cadmium acetate showed the highest inhibitory effects, with minimal inhibitory concentration ranging from 4-64 μg/ml and 16-256 μg/ml, respectively. The mean geometric for minimal fungicidal concentrations were, respectively, 26 μg/ml and 111.43 μg/ml for copper acetate and cadmium acetate. These results suggest that copper and cadmium can inhibit P. insidiosum growth, highlighting the greater inhibitory activity of copper acetate. In addition, our results propose that copper and/or cadmium compounds can be used in upcoming researches to formulate effective new complexed drugs against P. insidiosum in in vitro and in vivo experimental models., (© The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objective: To investigate the effects of rapid anesthesia and long-term sedation with the essential oils (EOs) of Myrcia sylvatica (EOMS) and Curcuma longa (EOCL) on biochemical and oxidative parameters in matrinxã., Study Design: Prospective, randomized, laboratory experiment., Animals: A total of 72 matrinxã (Brycon amazonicus) adults weighing 404.8 ± 27.9 g were divided into eight groups of nine fish., Methods: Biochemical and oxidative effects were investigated in plasma and tissues of matrinxã subjected to rapid anesthesia (5 minutes) or long-term sedation (360 minutes, simulating the practice of transport) with EOMS (200 μL L -1 and 10 μL L -1 , respectively) and EOCL (500 μL L -1 and 40 μL L -1 , respectively)., Results: Transport simulation without sedation or anesthesia increased lipid peroxidation levels in the gills and kidney of fish in the control group. Anesthesia and sedation with EOs decreased cortisol concentrations and increased lactate concentrations compared with controls. Lipid peroxidation was lower in the brain, gills, liver and kidney of sedated and anesthetized fish, than in the control group. Anesthesia with EOs increased the activity of superoxide dismutase and glutathione-S-transferase in the brain, and catalase in the liver and gills, compared with controls. Long-term sedation with EOs increased superoxide dismutase, glutathione peroxidase and glutathione reductase activities in the brain, catalase in the liver, glutathione peroxidase and glutathione reductase in the gills and superoxide dismutase in the kidney. In general, nonprotein thiols content and total reactive antioxidant potential of tissues were higher after anesthesia and sedation with EOs compared with the control group., Conclusions and Clinical Relevance: The concentrations of EOMS and EOCL used were effective at preventing a stress response and excess of reactive oxygen species formation. For these reasons, these substances may be recommended for use in the transportation of fish to improve survival and animal welfare., (Copyright © 2017 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

The aim of this study was to evaluate the activity of purinergic enzymes in lymphocytes and cardiac tissue of mice experimentally infected by Trypanosoma cruzi. Twelve female mice were used, divided into two groups (n = 6): uninfected and infected. On day 12 post-infection (PI), the animals were anesthetized and after euthanized, and samples were collected for analyses. Infected mice showed reduction in erythrocyte counts, hematocrit and hemoglobin concentration, as well as reduced number of total leukocytes in consequence of neutrophilia (P < 0.01). The number of monocytes increased in infected mice (P < 0.001), however the number of lymphocytes and eosinophils did not differ between groups (P > 0.05). The E-NTPDase (ATP and ADP substrate) and E-ADA activities in lymphocytes increased significantly in mice infected by T. cruzi (P < 0.01). In the heart, multiple pseudocysts containing amastigotes within cardiomyocytes were observed, as well as focally extensive severe necrosis associated with diffuse moderate to severe inflammatory infiltrate of lymphocytes. Although, the NTPDase activity (ATP and ADP substrate) in the cardiac homogenate did not differ between groups, a reduction on 5'-nucleotidase activity (P < 0.001) and an increase in the ADA activity in infected animals (P < 0.05) were observed. Thus, animals infected by T. cruzi experienced the disease, i.e., showed anemia, leucopenia, and heart lesions. Associated with this, purinergic enzymes showed altered activities, which might be related to the modulation of the inflammatory response., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Linalool is the main compound of many essential oils and occurs in two isomeric forms: S-(+)- and R-(-)-linalool. This study aimed to determine if linalool isomers have different antimicrobial and anesthetic properties in fish. For this purpose, these compounds were previously isolated from Lippia alba (Mill.)N. E. Brown and Ocimum americanum L. essential oils. Antimicrobial effects were evaluated through the microdilution test against Aeromonas hydrophila, an important fish disease etiologic agent. Induction time until sedation, anesthesia and recovery time were determined in silver catfish (Rhamdia quelen) through bath exposure (60, 180, 300 or 500 μL L-1). The results showed different biological properties for the isomers being S-(+)-linalool the only active against A. hydrophila at 3.2 mg mL-1. The sedation was induced without differences between the compounds, however R-(-)-linalool promoted faster anesthesia. There were no differences regarding the recovery time of the animals exposed to the linalool isomers. Although both S-(+)- and R-(-)-linalool can be used for sedative purposes, their use in A. hydrophila infection is inadvisable due to the high effective concentration. Considering anesthesia as the main objective, the R-(-)-linalool demonstrated clear advantages at lower concentration.

Gastrointestinal parasites are one of the biggest health problems faced in sheep, mainly due to their pathogenicity and resistance to drugs used to control these parasites. Thus, the following study aimed to assess the anthelmintic efficacy of Melaleuca alternifolia against Haemonchus contortus in gerbils (Meriones unguiculatus) experimentally infected. Three treatments were tested: M. alternifolia essential oil, popularly known as tea tree oil (TTO), a solid lipid nanocarrier made with essential oil of Melaleuca (nanoTTO), and terpinen-4-ol (terp-4-ol). In vivo studies were performed by determining the mean worm burden of H. contortus in gerbils TTO (0.75 mL/kg); nanoTTO (0.5 mL/kg) and terp-4-ol (0.5 mL l/kg) were able to reduce 46.36%; 48.64%, and 43.18% worm burden, respectively. H. contortus increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, as demonstrated by liver injury. It was found that the TTO, nanoTTO, and terp-4-ol were not toxic to liver and kidneys since hepatic and renal functions were not affected. Moreover, terp-4-ol was able to prevent increased levels of seric AST and ALT in infected animals, indicating a hepatoprotective effect. Thus, our results indicate that TTO, nanoTTO, and terp-4-ol are safe and efficient against H. contortus infection in gerbils, and possibly the terp-4-ol may be considered the compound present in the Melaleuca alternifolia responsible for parasitic action against H. contortus., (Copyright © 2016. Published by Elsevier Inc.)

Unlabelled: The bactericidal activity (minimum inhibitory concentration (MIC)-test) of Ocimum americanum (inflorescences) essential oil (OAEO) against Aeromonas hydrophila was determined in this study. Also investigated was the potential of OAEO and the main compound found in the oil (linalool) at subinhibitory concentrations to be inhibitors of haemolysis caused by Aer. hydrophila in fish erythrocytes. An in vivo experiment was conducted to evaluate the survival of fish (Rhamdia quelen) experimentally infected with Aer. hydrophila and exposed to OAEO. A second experiment was conducted to evaluate the in vitro and in vivo activity of OAEO (mix from inflorescences and leaves) against the parasite Gyrodactylus sp. The OAEO showed weak in vitro activity against Aer. hydrophila (6400 μg ml(-1) ). Subinhibitory concentrations of OAEO (100 μg ml(-1) ) inhibited haemolysis (90%) caused by Aer. hydrophila in fish erythrocytes, however, linalool did not inhibit haemolysis activity. At the low concentrations (10 and 20 mg l(-1) ) added to the water, OAEO promoted the survival of fish experimentally infected with Aer. hydrophila. Lastly, the OAEO mix (50 mg l(-1) ) was effective against Gyrodactylus sp., significantly reducing (60%) the number of parasites in the fish., Significance and Impact of the Study: Phytochemicals, such as essential oils (EOs) are a great source of new molecules and have shown potential to be used in aquaculture systems. However, additional studies focused on the in vivo efficacy, mode of action and identification of the active compounds are needed. This study determined the potential of Ocimum americanum EO for use against two important fish pathogens, Aeromonas hydrophila and Gyrodactylus sp., as well as providing preliminary information about the role of the main EO compound (linalool) against Aer. hydrophila virulence., (© 2016 The Society for Applied Microbiology.)

Parasitic diseases reflect the health and balance of ecosystems, affecting not only individuals but also entire populations or communities. The aim of this study was to report on the diversity of parasitic helminths detected in the feces of a wild feline in southern Brazil. Parasites were obtained from fecal samples, and four techniques were used for parasitological examination: direct examination, centrifugal flotation with zinc sulfate (Faust technique), simple sedimentation (Hoffman technique) and Baermann-Moraes. The parasites were identified through micrometry and morphology, as follows: Ancylostoma sp., Toxocara sp., Trichuridae, Aelurostrongylus abstrusus, Alaria sp., and Spirometra sp. We recorded the genus Ancylostoma parasitizing L. colocolo for the first time.

Imazapyr (IMY) and imazapic (IMI) are imidazolinone herbicides which have been associated in a commercial formulation (Kifix(®)). To date, there are no studies on the toxicity of an IMY+IMI herbicide in fish. This work aimed to assess the acute toxicity (24 and 96 h) of IMY+IMI (0, 0.488 and 4.88 µg/L) towards Rhamdia quelen through hematological, biochemical, immunological, ionoregulatory and enzymatic indexes. Red blood cell count was lower at 4.88 than at 0.488 µg/L (24 and 96 h); mean corpuscular volume was lower than control at both concentrations (24 h) and at 0.488 µg/L (96 h); lymphocytes declined at 4.88 µg/L comparing to control (96 h); and monocytes increased at 4.88 µg/L (96 h) in comparison with the respective control and with 4.88 µg/L at 24h. Aspartate aminotransferase was higher at 0.488 µg/L (96 h) than the respective control and the respective concentration at 24 h; uric acid reduced at 4.88 µg/L comparing with 0.488 µg/L (96 h); and cortisol was lower at 4.88 µg/L compared to 0.488 µg/L and control (96 h). Herbicide exposure lowered plasma bactericidal activity at both concentrations (24 h) and at 0.488 µg/L (96 h); and plasma complement activity declined at 4.88 µg/L comparing with 0.488 µg/L and control (96 h), and was lower at all concentrations at 96 h than at 24 h. Plasma K(+) levels were higher at 4.88µg/L than in the remaining groups (24 and 96h); and Na(+) levels decreased at 4.88 µg/L compared to control (96 h). Na(+)/K(+)-ATPase and H(+)-ATPase activities in gills were lower at 4.88 µg/L comparing with control (24 h) and with the respective concentration at 96 h; and AChE activity in brain was higher at 0.488 and 4.88 µg/L than control (24 h) and the respective concentrations at 96 h, while in muscle it was higher at 0.488 and 4.88 µg/L than control (96 h) and the respective concentrations at 24 h. The present findings demonstrate that, despite IMY+IMI targets the animal-absent AHAS enzyme, such formulation displayed an acute toxic effect upon R. quelen homeostasis by impacting on vital functions such as immune defense, metabolism, ionoregulation and neurotransmission., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Rhodococcus equi preferentially infects macrophages causing pyogranulomatous pneumonia in young foals. Both the vapA and rhbC genes are up-regulated in an iron (Fe)-deprived environment, such as that found within macrophages. Chloroquine (CQ) is a drug widely used against malaria that suppresses the intracellular availability of Fe in eukaryotic cells. The main objective of this study was to evaluate the ability of CQ to inhibit replication of virulent R. equi within murine (J774A.1) and foal alveolar macrophages (AMs) and to verify whether the mechanism of inhibition could be Fe-deprivation-dependent. CQ effect on R. equi extracellular survival and toxicity to J774A.1 were evaluated. R. equi survival within J774A.1 and foal AMs was evaluated under CQ (10 and 20μM), bovine saturated transferrin (bHTF), and bovine unsaturated transferrin (bATF) exposure. To explore the action mechanism of CQ, the superoxide anion production, the lysozyme activity, as well as the relative mRNA expression of vapA and rhbC were examined. CQ at≤20μM had no effect on R. equi extracellular multiplication and J774A.1 viability. Exposure to CQ significantly and markedly reduced survival of R. equi within J774A.1 and foal AMs. Treatment with bHTF did not reverse CQ effect on R. equi. Exposure to CQ did not affected superoxide anion production or lysozyme activity, however vapA and rhbC expression was significantly increased. Our results reinforce the hypothesis that intracellular availability of Fe is required for R. equi survival, and our initial hypothesis that CQ can limit replication of R. equi in J774A.1 and foal AMs, most likely by Fe starvation., (Copyright © 2016 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the susceptibility in vitro and in vivo of Trypanosoma evansi to terpinen-4-ol, γ-terpinene and α-terpinene, the three main compounds of tea tree oil (Melaleuca alternifolia) with known efficacy in the treatment of trypanosomosis. In vitro, a trypanocidal effect of terpinen-4-ol, γ-terpinene, and α-terpinene was observed when used alone or associated at 0.5, 1 and 2% concentrations i.e., the α-terpinene showed a faster trypanocidal effect when compared to chemotherapy (diminazene aceturate - D.A.). In vivo studies were performed in two experiments: I and II where experiment I used T. evansi infected mice treated with terpinen-4-ol, γ-terpinene and α-terpinene alone (at a dose of 1.0 mL kg(-1)) or associated (two compounds, dose of 0.5 mL kg(-1) of each compound; tree compounds, dose of 0.335 mL kg(-1) of each compound); Treatment with α-terpinene was able to extend animal longevity, but showed no curative efficacy. In experiment II, T. evansi infected mice were treated with D.A. associate with α-terpinene, where a curative efficacy of 57.14% was found, a much better result when D.A. was used alone (14.28%). In summary, α-terpinene associated with D.A. can be used as an alternative treatment for T. evansi infection. The compound α-terpinene from M. alternifolia essential oil is the one responsible for the trypanocidal effect, a fact confirmed by in vitro results and the increased longevity observed on treated mice., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The aim of this study was to estimate the prevalence of equine strangles and to identify associated risk factors for this disease through a cross-sectional study of nasal swabs. Nasal swabs (n=1010) from healthy equines (absence of nasal discharge, lymphadenopathy and cough) from 341 farms were plated on 5% blood agar; of these horses, 24 were identified as positive for Streptococcus equi through isolation, PCR and DNA sequencing. The estimated prevalence for individual animals was 2.3%, and for herds, it was 5.86%. Statistical analysis identified the following as associated risk factors: the number of group events that were attended by the equines (PR: 1.06); the sharing of food containers (PR: 3.74); and at least one previous positive diagnosis of strangles on the farm (PR: 3.20). These results constitute an epidemiological contribution to the horse industry and may support measures for the future control of the disease., (Copyright © 2015. Published by Elsevier Ltd.)

The effects of adding rutin to the diet (0, 0.15 or 0.30%) of silver catfish for 21 days on blood parameters, oxidative stress biomarkers and pituitary hormones expression were investigated. Fish that received the diet containing 0.15% rutin exhibited reduced plasma cortisol levels. The levels of lipid peroxidation were lowered in the all tissues of animals receiving the diet containing rutin. Rutin increased the activity of the superoxide dismutase (SOD), catalase (CAT), nonprotein thiols (NPSH), ascorbic acid content (AA) and total reactive antioxidant potential (TRAP) in the brain; glutathione S-transferase (GST) activity and TRAP in the gills; SOD, CAT and GST activity, NPSH, AA levels and TRAP in the liver; CAT and GST activity and TRAP levels in the kidneys; and glutathione peroxidase activity, NPSH, AA levels and TRAP in the muscle. There were no changes regarding the expression of growth hormone, prolactin and somatolactin in fish fed with the diet containing rutin when compared with the control. The supplementation of rutin to the diet of fish is beneficial because it increases the antioxidant responses of tissues.

(+)-Dehydrofukinone (DHF) is a major component of the essential oil of Nectandra grandiflora (Lauraceae), and exerts a depressant effect on the central nervous system of fish. However, the neuronal mechanism underlying DHF action remains unknown. This study aimed to investigate the action of DHF on GABAA receptors using a silver catfish (Rhamdia quelen) model. Additionally, we investigated the effect of DHF exposure on stress-induced cortisol modulation. Chemical identification was performed using gas chromatography-mass spectrometry and purity was evaluated using gas chromatography with a flame ionization detector. To an aquarium, we applied between 2.5 and 50 mg/L DHF diluted in ethanol, in combination with 42.7 mg/L diazepam. DHF within the range of 10-20 mg/L acted collaboratively in combination with diazepam, but the sedative action of DHF was reversed by 3 mg/L flumazenil. Additionally, fish exposed for 24 h to 2.5-20 mg/L DHF showed no side effects and there was sustained sedation during the first 12 h of drug exposure with 10-20 mg/L DHF. DHF pretreatment did not increase plasma cortisol levels in fish subjected to a stress protocol. Moreover, the stress-induced cortisol peak was absent following pretreatment with 20 mg/L DHF. DHF proved to be a relatively safe sedative or anesthetic, which interacts with GABAergic and cortisol pathways in fish.