Antimicrobial activity of cationic water-soluble porphyrin against multidrug-resistant bacteria in biofilms and canine skin samples.

Multidrug-resistant (MDR) microorganisms pose a threat to animal health, particularly in integumentary diseases, which can be caused by multiple organisms and often manifest as biofilms, hindering treatment effectiveness. We evaluated the antimicrobial activity of antimicrobial photodynamic therapy (aPDT) using a water-soluble tetra-cationic porphyrin (4-H2TMeP) against MDR bacteria cultured in biofilm and in mono and polyculture grown on canine skin samples. We utilized 4-H2TMeP porphyrin against MDR Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus pseudintermedius. A non-cytotoxic concentration of 4-H2TMeP (40 µM), previously shown to be effective in vitro against these bacteria cultured in solution, was employed. Biofilms were treated with 4-H2TMeP and subjected to light irradiation for 30, 60, and 90 min. Monocultures on canine skin samples were treated with 4-H2TMeP and irradiated for 30 (S. pseudintermedius), 60 (E. coli), or 60 and 90 min (P. aeruginosa). Polycultures of S. pseudintermedius and E. coli were treated with light for 60 and 90 min. The efficacy of aPDT was evaluated by plating light-exposed biofilms, mono and polycultures of bacteria obtained from skin samples exposed to light and kept in the dark. Colony-forming units were counted after 24 h of incubation at 37 °C. aPDT using 4-H2TMeP reduced bacterial concentrations of S. pseudintermedius and E. coli biofilms. Additionally, it significantly reduced bacterial concentrations cultivated on skin samples, with a particular emphasis on S. pseudintermedius. These findings indicate that aPDT with 4-H2TMeP is a promising alternative treatment against MDR bacteria in animal skin infections and should be further explored through in vivo research.

In vitro antimicrobial, antibiofilm photodynamic activity, and molecular dynamic simulations of tetra-cationic porphyrinmembrane interactions against foodborne microorganisms.

This manuscript presents a new report on the in vitro antimicrobial photo-inactivation of foodborne microorganisms (Salmonella spp. and Listeria monocytogenes) using tetra-cationic porphyrins. Isomeric tetra-cationic porphyrins (3MeTPyP, 4MeTPyP, 3PtTPyP, and 4PtTPyP) were tested, and antimicrobial activity assays were performed at specific photosensitizer concentrations under dark and white-light LED irradiation conditions. Among the tested bacterial strains, 4MeTPyP exhibited the highest efficiency, inhibiting bacterial growth within just 60 min at low concentrations (17.5 µM). The minimal inhibitory concentration of 4MeTPyP increased when reactive oxygen species scavengers were present, indicating the significant involvement of singlet oxygen species in the photooxidation mechanism. Furthermore, the checkerboard assay testing the association of 4MeTPyP showed an indifferent effect. Atomic force microscopy analyses and dynamic simulations were conducted to enhance our understanding of the interaction between this porphyrin and the strain's membrane.

Economic losses caused by mastitis and the influence of climate variation on the occurrence of the disease in a dairy cattle farm in southern Brazil.

This study evaluated the economic impacts caused by mastitis in a small dairy farm with similar characteristics and production to most dairy farms in southern Brazil and investigated if climatic variations influenced mastitis occurrence in the region. A farm with, on average, 45 lactating Holstein cattle was monitored from November 2021 to October 2022, and data on mastitis cases, bulk tank milk somatic cell count, animal treatment costs, milk production, animal disposal costs, and production losses were collected. Monthly averages of temperature, relative humidity (RH), and rainfall in the region were obtained. The greatest loss was related to the drop in milk production, resulting in 63.8% of total losses, followed by animal disposal (29.5%), milk disposal (4.6%), and treating animals with mastitis (2.0%), totaling a 10.6% reduction in the annual gross income. There were negative correlations between the clinical mastitis rate and monthly RH and between subclinical mastitis and temperature; the occurrence of subclinical mastitis and average RH were positively correlated. Our findings showed that mastitis negatively impacted the economy and that climate influenced mastitis occurrence.

Antibiofilm action using water-soluble tetra-cationic porphyrin and antibacterial photodynamic therapy against Moraxella spp. from cattle.

Infectious bovine keratoconjunctivitis (IBK) is the most important eye disease in ruminants worldwide. Moraxella bovis and Moraxella bovoculi can form biofilm and are frequently isolated from affected animals. Antimicrobials are used worldwide to treat clinical cases of IBK, although they have limited success in clearing the infection. Therefore, photodynamic therapy using porphyrins as photosensitizing molecules is an alternative method to eliminate microorganisms, including biofilms. We evaluated the antibacterial activity of a zinc(II) metalloporphyrin (ZnTMeP) against M. bovis and M. bovoculi biofilms since this compound can efficiently inactivate planktonic Moraxella spp. This study was carried out with two reference strains of Moraxella spp. (M. bovis: ATCC® 10900 and M. bovoculli: ATCC® BAA1259). The antibacterial activity of 4.0 µM of the ZnTMeP porphyrin was evaluated on forming and consolidate biofilms with three 30-min cycles of white-light exposure for three days. The ZnTMeP porphyrin reduced M. bovis and M. bovoculi biofilm formation. In addition, ZnTMeP partially destroyed consolidated M. bovoculi biofilms in the second white-light irradiation cycle, although the porphyrin had no effect against the consolidated biofilm of M. bovis. Despite the biofilm still not being completely inactivated, our findings are promising and encourage further experiments using the phototherapy protocol.

Increased seroprevalence of anti-Toxoplasma gondii antibodies in dogs in southern Brazil after an outbreak of human toxoplasmosis.

Toxoplasmosis is a worldwide disease caused by Toxoplasma gondii, which can infect diverse hosts, including dogs. Although T. gondii infection in dogs is usually subclinical, they are susceptible to infection and develop a specific immune response to the parasite. In 2018, the largest outbreak of human toxoplasmosis in the world occurred in Santa Maria, in southern Brazil; however, the impact of this outbreak on other hosts was not investigated at the time. Considering that dogs often share the same environmental sources of infection as humans, mainly water sources, and that in Brazil, the detection rates of anti-T. gondii immunoglobulin G (IgG) in dogs is very high, this study investigated the frequency of anti-T. gondii IgG in dogs in Santa Maria before and after the outbreak. A total of 2.245 serum samples were analyzed, 1159 collected before the outbreak and 1086 collected after the outbreak. Serum samples were tested for anti-T. gondii antibodies using an indirect immunofluorescence antibody test (IFAT). The infection detection of T. gondii was 16% (185/1159) before the outbreak and 43% (466/1086) after the outbreak. These results showed the infection of dogs with T. gondii and the high frequency of anti-T. gondii antibodies in dogs after the outbreak in humans in 2018, reinforcing water as a possible source of infection and the importance of including toxoplasmosis in the differential diagnosis of dogs.

Five years of the biggest outbreak of human toxoplasmosis in Santa Maria, Brazil: a review.

Toxoplasmosis is a parasitic disease caused by infection with the protozoan Toxoplasma gondii. In 2018, the first cases of people with clinical signs of acute febrile syndrome were reported, and in the same year, the largest outbreak of human toxoplasmosis ever described in the literature was reported. In this sense, the present work sought to describe the evolution of the outbreak cases in the municipality of Santa Maria, Rio Grande do Sul State, Brazil, as well as the studies conducted and published during and after the outbreak in the municipality (the period between 2018 and 2023). In addition, the discussion of public policies and their modifications after the notification of this outbreak. As a result of this research, verifying the evolution of notified and confirmed cases, the possibility of detection and genotypic characterization of T. gondii and the possibility of co-infections and evaluation of the humoral response is possible. With regard to public policies, the importance of detecting the agent through the heel prick test and increasing the monitoring of water quality to prevent outbreaks.

Increased frequency of detection of anti-Toxoplasma gondii antibodies in domestic cats after outbreak of human toxoplasmosis.

Toxoplasmosis affects various organisms, including humans. In 2018, the largest outbreak of human toxoplasmosis described so far was reported in southern Brazil, with 809 human cases reported, and water as the potentially primary source of infection. Therefore, in this study, we aimed to evaluate the seroprevalence of Toxoplasma gondii in naturally infected domestic cats before and after the human toxoplasmosis outbreak, as well as the potential for environmental contamination by the number of cats infected after the outbreak. We evaluated 381 serum samples from domestic cats in southern Brazil, using an indirect immunofluorescence assay, with samples considered positive at a titer of 1:20. We found that 73% (204/279) and 27% (75/279) of the samples analyzed before the outbreak were negative and positive, respectively. After the outbreak, 62% (69/112) were negative of the samples were and 38% (43/112) were positive. Notably, the proportion of positive samples before the outbreak before (27%) was significantly lower than that after the outbreak (38%; P = 0.020). Therefore, the increased seroprevalence of T. gondii in cats was probably correlated with the ingestion of contaminated water. Therefore, it is important to monitor animals, mainly definitive hosts, after toxoplasmosis outbreaks, considering that these animals can contaminate the environment and, consequently, humans.

Characterization of novel of temperate phages of Staphylococcus aureus isolated from bovine milk.

Bovine mastitis is an important disease in dairy cows, and Staphylococcus aureus is the most prevalent microorganism. Bacteriophages are considered an alternative to treat bacterial infections due to antimicrobial resistance crisis. In this study, we isolated and characterized novel S. aureus temperate phages, namely B_UFSM4 and B_UFSM5, from bovine milk. The complete genomes of B_UFSM4 and B_UFSM5 have 41.396 bp and 41.829 bp, respectively. The viruses have double-stranded DNA and linear architecture. Phylogenic similarity was observed by proteome with Staphylococcus phage phiPV83, CN125 and JS01. Therefore, the phages were classified into the family Siphoviridae, genus Biseptimavirus and order Caudovirales. In the host range, the B_UFSM4 and B_UFSM5 had lytic activity of 45.8% and 54.16%, respectively, inclusive on isolates from Staphylococcus sciuri and Rothia terrae. Thus, in this study, species novel of S. aureus temperate phages was isolated and characterized, these phages reveal similarities to each other; however, they are distinct from other species of S. aureus phages of the family Siphoviridae.

Safety and efficacy of Lucilia cuprina maggots on treating an induced infected wound in Wistar rats.

Infection is one of the main complications that hinder wound healing. Currently, antibiotic-resistant bacteria, such as Methicilin-resistant Staphylococcus aureus (MRSA), are a concern worldwide for both humans and animals. Maggot therapy is re-emerging as an alternative to intractable wounds and may be an option to the traditional antibiotic treatment. Although the species of choice is Lucilia sericata, reports of clinical use have led us to evaluate the efficacy and safety of using Lucilia cuprina larvae on induced infected wounds in Wistar rats. In short, 32 male Wistar rats were divided into 4 groups: Group I - saline solution treated; Group II - antibiotic-treated; Group III - treated with larval debridement, and Group IV - without wound and treatment. Skin wounds were induced in groups I, II and III. All treatments were performed once and held for 48 h. Clinical, microbiological, histopathological, hematological, and biochemical analyses were done. Significant wound area contraction was found (>95%) in group III on day 9 compared to day 15 in group II. Complete elimination (0.0 ± 0.0 CFU/mL) of bioburden was achieved after the second treatment (day 6) in both the II and III groups, compared to an increase in Group I (6.51 ± 0.37 CFU/mL). A cleaner wound was also observed in the histopathological evaluation of group III, with adequate collagen formation and re-epithelialization on day 15. Furthermore, larvae increased blood platelet levels after the first treatment. L. cuprina larvae have proven safe and effective in accelerating wound treatment and eliminating MRSA.

Dynamics of Leishmania spp. infection in dogs from an unaffected region in transition to a visceral leishmaniasis transmission area, Rio Grande do Sul, Brazil.

In Brazil, visceral leishmaniasis (VL) has been expanding and urbanizing, mainly in non-endemic areas such as the State of Rio Grande do Sul. Considering that infected dogs are the main reservoirs of VL in urban areas, the present study aimed to evaluate the propagation of canine visceral leishmaniasis (CVL) infection from an unaffected region in transition to a VL transmission area. For this, 1159 and 1087 samples of canine serum from 2015 and 2021, respectively, were analyzed, using the indirect immunofluorescence test. In addition, necropsy reports between 2007 and 2021 were evaluated. The results showed a prevalence of anti-Leishmania spp. antibodies of 7.5% in the samples from 2015, while in 2021 samples, it was 23.5%, with an incidence of 0.4 cases per 100 dogs. It is noteworthy that in 2007, there was no record of CVL as the cause of death in the pathological reports, and in 2021, 41 diagnoses were made with the protozoan being a determinant of the death of the animal. These values indicate an increasing trend in the prevalence and incidence coefficients of CVL. The results of this study allowed us to verify the spread of the disease from an unaffected region to a transmission area of the agent, as well as provide subsidies for health authorities to implement improvements in the CVL control program in the municipality, to mitigate the emergence of human cases of the disease.

Prevalence and molecular detection of Leishmania spp. in bats from Rio Grande do Sul state, Brazil.

This study aimed to detect the occurrence of infection by Leishmania spp.in bats from 34 municipalities of Rio Grande do Sul state (RS; southern Brazil) from 2016 to 2021. A total of 109 bats were provided by the Centro Estadual de Vigilância em Saúde of RS, including six species belonged to Molossidae family, six to Vespertilionidae family, and two to Phyllostomidae family. Leishmania spp. was identified using the nested-PCR method by amplifying the SSU rDNA ribosomal subunit gene into four organ pools: (1) the liver, spleen, and lymph node; (2) heart and lungs; (3) skin; and (4) bone marrow of each bat. Three (3/109, 2.7%) animals tested positive for Leishmania spp. The respective PCR-positive organs came from pools 1 and 3. Two bats (Tadarida brasiliensis) were from the municipality of Canoas, and sequences analysis confirms the species identification as Leishmania infantum. In the third bat (Molossus molossus), from Rio Grande, it was not possible to determine the protozoa species, being considered Leishmania spp. Our results indicate that bats can participate in the biological cycle of Leishmania spp. and perform as host, reservoir, and/or source of infection of the protozoa in different areas of RS. More studies will be needed to elucidate the role of these Chiropteras in the circulation of Leishmania spp. This is the first study reporting the occurrence of Leishmania spp. in bats in Rio Grande do Sul state, southern Brazil.

Acaricide activity of organo-modified siloxane and acaricide associations in Rhipicephalus microplus.

Search to a new alternative to control bovine ticks (Rhipicephalus microplus), the present study aimed to evaluate the acaricidal activity of organo-modified siloxane alone and in association with different commercial products or with piperonyl butoxide (BPO). Engorged females were subjected to an in vitro immersion test and 10 groups were used: control, 0.5% siloxane, 1% siloxane, 2% siloxane, 0.5% siloxane + 5% BPO, 1% siloxane + 5% BPO, 2% siloxane + 5% BPO, commercial product, 0.5% siloxane + commercial product, and 1% siloxane + commercial product. After immersion, engorged females were incubated for 14 days for oviposition and hatchability tests. Another immersion test was performed with 5% siloxane and 2.5% siloxane + 10% BPO to evaluate the histopathological changes. Then, engorged females were incubated for 0, 2, 4, 8, 12, 24, 36, 48, 60, 72, 84, 96, 120, 144, and 168 h and immersed in 10% formaldehyde for later analyses. The unassociated siloxane showed an acaricide efficacy of 93.88% at 2% concentration, and when associated with 5% BPO, it reached 100% efficacy at all tested concentrations. The tested commercial products showed enhanced efficacy when associated with siloxane. Histopathological analysis showed cell changes in both treatments and total cell disintegration after 120 h in the 5% siloxane group and after 96 h in the 2.5% siloxane + 10% BPO group. Therefore, siloxane alone or in combination is an alternative against R. microplus, and siloxane enhances the efficacy of available commercial products.

GAPDH, rhbC, and vapA gene expression in Rhodococcus equi cultured under different iron concentrations.

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.

In vitro algicidal effect of polypyrrole on Prototheca species isolates from bovine mastitisAlgicidal activity of polypyrrole on Prototheca spp.

Algae of the genus Prototheca are microorganisms involved in the occurrence of diseases in humans and animals. In bovine species, Prototheca spp. cause environmental mastitis, productive losses in dairy herds, mainly leading to the discard of infected cows. Currently, there are no effective anti-Prototheca spp. drugs to combat this infection. Thus, the search for an efficacious therapy for Prototheca spp. infections have become essential. Highly soluble polypyrrole (Ppy) is a molecule with known antimicrobial activity. This study aimed to characterize Prototheca spp. isolates from bovine mastitis as well as to evaluate the susceptibility profile and to verify the morphological alterations on Prototheca spp. isolates treated with Ppy. In this research, 36 Brazilian isolates of Prototheca spp. were characterized by restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR) assay for the mitochondrial cytB gene. Additionally, Ppy algicidal activity against these isolates of Prototheca spp. was assessed by minimal microbicidal concentration method in microplates. Further, scanning electron microscopy (SEM) was performed in order to verify the morphological alterations on Prototheca spp. isolates in response to Ppy. The isolates were characterized as belonging to Prototheca zopfii genotype 2 (35/36) and Prototheca blaschkeae (1/36). Ppy had an algicidal effect on all isolates tested at concentrations ranging from 15.625 µg ml-1 to 62.5 µg ml-1. SEM showed changes on planktonic and sessile P. zopfii, including a decrease of the number of cells with the presence of an amorphous substance involving the cells. The algicidal activity of Ppy suggests the therapeutic potential of this molecule in the prevention and treatment of Prototheca spp. in bovine mastitis.

Aeromonas hydrophila infection in silver catfish causes hyperlocomotion related to stress.

Aeromonosis is a fish disease that leads to haemorrhagic septicaemia and high mortality. The detection of early behavioural changes associated to this disease could be helpful in anticipating the initiation of treatment, increasing the probability of success. The influence of this disease on the hypothalamic-pituitary-interrenal (HPI) axis and on the brain expression of heat shock proteins (HSP) is little known. Therefore, the aim of this study was to evaluate the effect of Aeromonas hydrophila infection on individual behaviour and brain expression of genes related to stress (slc6a2, hsp90, hspa12a, hsd20b, hsd11b2, crh) in silver catfish (Rhamdia quelen). Thirty fish were divided into healthy and infected groups. The fish of the infected group were inoculated intramuscularly with 50 µL of bacterial suspension (6.4 × 108 CFU/mL), while control animals received 50 µL of saline. On day five post-infection, animals were submitted to the novel tank test, euthanized, and the brain was collected for molecular analysis. Infected fish swam more in the unknown aquarium and presented an increase in brain expression of genes related to HSP (hspa12a) and the route of cortisol synthesis (crh) when compared to uninfected fish. Therefore, this disease causes hyperlocomotion related to stress.

New variants of canine parvovirus in dogs in southern Brazil.

Carnivore protoparvovirus 1 (canine parvovirus 2, CPV-2) has undergone a rapid evolution through mutations in the capsid protein VP2, giving rise to variants associated with unique clinicopathological and immunological features. VP2 is a major capsid protein involved in key steps of virus biology, including interactions with cellular receptors and with the immune system. This study analyzed the complete VP2 coding sequence of 38 CPV-2 isolates obtained from dogs with clinical parvovirosis in southern Brazil. Amplicons encompassing the whole VP2 coding region were subjected to nucleotide sequencing, and predicted amino acid sequences were analyzed to identify molecular markers of viral variants. Viral variants were classified as CPV-2a, -2b or -2c based on the presence of the amino acid Asn, Asp or Glu, respectively, at VP2 residue 426. Amino acid sequence analysis identified 20 CPV-2c and four CPV-2b isolates. Eleven viruses were identified as New CPV-2a, two as New CPV-2b, and one resembled the original CPV-2 and was designated CPV-2-like. In addition to the mutation at amino acid 426 of VP2, new 2a/2b variants containing a Ser297Ala mutation at residue 297 were identified. CPV-2-like samples contained some mutations that were also present in the original CPV-2 isolate, including as Leu, Thr, Ala and Asp at residues 87, 101, 300 and 305, respectively. The New CPV-2a isolates had three additional mutations (Phe267Tyr, Tyr324Ile and Thr440Ala) associated with selective pressure and development of disease in vaccinated dogs. The resemblance of the CPV-2-like isolate to CPV-2 suggests reemergence of CPV-2 and/or evolution from vaccine strains. Phylogenetic analysis grouped the variants with their respective reference strains, in general, according to amino acid changes. These results demonstrate the high VP2 diversity of CPV circulating in dogs in southern Brazil and indicate the emergence of new viral variants that differ markedly from the current vaccine strains.

Molecular identification of Sarcocystis species in wild boar (Sus scrofa) and pigs (Sus scrofa domesticus) in Brazil.

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.

Molecular identification of Sarcocystis neurona in tissues of wild boars (Sus scrofa) in the border region between Brazil and Uruguay.

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.

Molecular detection of DNA from Trypanosoma spp. and Leishmania spp. in wild boar (Sus scrofa) tissues.

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.