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Aim: To evaluate the urinary biomarkers related to sepsis in preterm newborns (NBs) and to investigate the predictive capacity of these biomarkers for a longer hospital stay. Methods: Serum and urine were collected from 27 healthy NBs, 24 NBs with neonatal infection without sepsis and 11 NBs with sepsis for the measurement of sindecan-1, lipocalin associated with urinary neutrophil gelatinase (uNGAL), urinary cystatin-C (uCysC) and urinary kidney injury molecule-1. Results: Levels of uNGAL and urinary cystatin-C were elevated in NBs with sepsis and neonatal infection, and uNGAL was significant predictor of hospital stay longer than 30 days (odds ratio: 1.052; 95% CI: 1.012-1.093; p  = 0.01). Conclusion: uNGAL was associated with sepsis in preterm NBs and was useful to predict extended hospital stay.

Substance use disorder (SUD) is a significant health problem, and trauma exposure is a known risk factor for the escalation of substance use. However, the shared neural mechanisms through which trauma is associated with substance use are still unknown. Therefore, we systematically review neuroimaging studies focusing on three domains that may contribute to the overlapping mechanisms of SUD and trauma-reward salience, negative emotionality, and inhibition. Using PRISMA guidelines, we identified 45 studies utilizing tasks measuring these domains in alcohol, tobacco, and cannabis use groups. Greater reward, lesser regulation of inhibitory processes, and mixed findings of negative emotionality processes in individuals who use substances versus controls were found. Specifically, greater orbitofrontal cortex, ventral tegmental area, striatum, amygdala, and hippocampal activation was found in response to reward-related tasks, and reduced activation was found in the inferior frontal gyrus and hippocampus in response to inhibition-related tasks. Importantly, no studies in trauma-exposed individuals met our review criteria. Future studies examining the role of trauma-related factors are needed, and more studies should explore inhibition- and negative-emotionality domains in individuals who use substances to uncover clinically significant alterations in these domains that place an individual at greater risk for developing a SUD., Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2024.)

Maintaining reliable energy supplies with resilience to extreme weather, water shortage and rising electricity and cooling demand is crucial to successfully implementing the clean energy transition. The integrated power and water systems found in several hyper-arid countries, featuring cooling-driven electrical demand and near-total dependence on seawater desalination, offer case studies illustrating energy system robustness to these conditions. We use linear optimization to minimize costs in a model system based on the resiliency-oriented energy system of the United Arab Emirates (UAE) while progressively decarbonizing the energy mix. We demonstrate that high levels of renewable energy adoption are economically favorable under conservative future technology cost assumptions, even including strict resiliency requirements, and draw conclusions for other energy systems making the transition to renewable energy under challenging climatic conditions, especially regarding the role of water desalination, demand flexibility, energy storage, and suitability of conventional design rules for ensuring resilience in renewable-dominated systems., Competing Interests: The authors declare no competing interests., (© 2024 The Authors.)

An allergy to bites from Culicoides (Diptera: Ceratopogonidae) occurs because of a hypersensitivity reaction caused by the inoculation of insect salivary antigens during the bite, resulting in immune-mediated dermatitis. To the best of our knowledge, no previous studies have focused on allergic dermatitis in donkeys in Brazil. Therefore, this study aimed to describe the epidemiological, clinicopathological, and therapeutic aspects of allergic dermatitis in donkeys and to identify the insects involved in its epidemiology. This study reported the occurrence of dermatitis in 17 animals. The clinical signs were restlessness and severe itching. Skin lesions were found on the head, depigmented areas of the muzzle and cheeks, flanks, pelvic and thoracic limbs, and the scrotal sac. The lesions were characterized by areas of alopecia with crusts accompanied by serosanguineous exudates. Histologically, the lesions were characterized as moderate superficial dermatitis with irregular epidermal acanthosis and pronounced diffuse orthokeratotic hyperkeratosis. In total, 378 Culicoides specimens were collected, with Culicoides ocumarensis Ortiz being the most abundant species. The combined application of copaiba oil and a multivitamin emulsion exhibited potential for topical treatment of allergic dermatitis caused by insect bites in donkeys. Our study revealed an association between allergic dermatitis in donkeys and Culicoides .

This is the first description of cutaneous mucormycosis in buffalo in the Brazilian Amazon biome. All buffalo showed apathy, inappetence, weight loss, reluctance to move, and prolonged sternal decubitus. Of the four affected animals, two died 15 and 30 days after the appearance of clinical signs. In the initial phase, the skin lesions were rounded areas with dry central regions, sensitive to palpation, with protruding edges and diameters ranging from 8 cm to 15 cm. These areas of necrosis were isolated or coalescing and present mainly on the limbs and sides. In an advanced stage of the disease, there was detachment of the skin from the necrotic areas with extensive wound formation, which sometimes exposed the subcutaneous tissue. The histopathology of the skin showed a multifocal inflammatory infiltrate composed of intact and degenerated eosinophils surrounded by epithelioid macrophages. At the center of these areas was a focally extensive area of epidermal ulceration characterized by intact and degenerated neutrophils, the necrosis of epithelial cells, and the accumulation of fibrin and erythrocytes. The mycological culture was positive for Rhizopus sp. The diagnosis of cutaneous dermatitis caused by Rhizopus sp. was based on clinical signs, macroscopic and histopathological findings, and the identification of the fungus by mycological and molecular techniques.

Bovine Dewclaw Dermatitis (BDCD) is a hoof disease characterized by inflammation of the second and fifth accessory digits and the skin in this region. This pathology is poorly described in the literature; however, it has recently been observed in beef cattle in the Amazon Biome, Brazil. The objective of this study was to perform a clinical diagnosis and identify the risk factors associated with BDCD onset in cattle in the studied biome. Samples were collected from eight farms with extensive breeding systems located in Xinguara, Rondon do Pará, Curionópolis, and Ipixuna do Pará in the state of Pará, Brazil. A total of 706 Nellore and Nellore crossbred with taurine bovine of both sexes were evaluated, with males aged between 2 and 4 years and a mean weight of 650 kg, and females aged between 2 and 11 years and a mean weight of 400 kg. Distal extremities were inspected during cattle management, and in cases of dewclaw lesions, a specific examination was carried out after proper restraint. Cattle were diagnosed with BDCD on all farms analyzed. Of the 706 cattle inspected, 49 (6.94%) showed BDCD, of which 19 (38.77%) were Nellore and 30 (61.22%) were crossbred. This was the first study to determine BDCD's occurrence in extensive farming systems in the Amazon region, also showing that pastures with large amounts of stumps and stones, the physical structure of pens, and trauma and injury incidence during animal management are the most important predisposing factors for the onset of BDCD.

Meningitis is the inflammation of the membranes surrounding the central nervous system and is poorly described in water buffaloes. Five cases of meningitis in adults buffaloes of the Murrah and Mediterranean breads were studied. All buffaloes came from a farm located in the municipality of Castanhal, Pará, Brazil at different times. Clinical examination showed neurological clinical signs, such as apathy, reluctance to move, spastic paresis especially of the pelvic limbs, hypermetria, difficulty getting up, pressing of the head into obstacles and convulsion. In three buffaloes, a large part of the horn had been lost, exposing the corresponding frontal sinus, through which a bloody to purulent exudate flowed. The hemogram revealed neutrophilic leukocytosis. At necropsy, adherence of the dura mater to the periosteum and a purulent to fibrinopurulent exudate were observed in the sulci of the cerebral cortex and on the pia mater over almost the entire surface of the brain and throughout the spinal cord. The cerebrospinal fluid had a cloudy aspect with fibrin filaments. The histopathology of buffaloes confirmed the diagnosis of bacterial fibrinopurulent meningitis. Buffaloes are susceptible to bacterial inflammation of the meninges due to fractures of the base of the horn and mostly present with neurological manifestations.

The epidemiological and clinical pathological aspects of polioencephalomalacia (PEM) in five buffaloes in the Amazon biome are described. Epidemiological data were obtained during the clinical visit in which buffalos were submitted to an examination of the nervous system. The diagnosis of PEM was based on epidemiological, clinical-pathological, and histopathological findings, similar to findings in other ruminants. In the clinical examination, all buffaloes had a body score between 2.5 and 3 (scale from 1 to 5), with decreased alertness, postural changes, marked hypermetria when stimulated to move, total or partial blindness demonstrated by colliding with the corral structures, head pressing and circumduction of the hindquarters when supported on the thoracic limbs, opisthotonos, muscle tremors, convulsions, paddling movements, sialorrhea, decrease in palpebral and pupillary reflexes, and eyeball rotation placing the pupillary slit in a vertical position. At necropsy, mild lesions characterized by edema and flattening of the cerebral convolutions were observed. A histopathological examination showed laminar necrosis of the cerebral cortex. The cause of PEM in the studied buffaloes was not established, which indicates the need for further studies to elucidate this disease in the species, particularly in the Amazon region.

Leucoderma is a condition that affects the skin and hair of animals, causing depigmentation and acromotrichia. In buffaloes, this condition results in significant economic losses for the production chain due to its impact on the leather trade. This study aimed to investigate the epidemiological and clinicopathological aspects of leucoderma in buffaloes in the Amazon biome and describe the prophylactic treatment to control the disease. The study included 40 buffaloes, 16 males and 24 females, aged between 1 and 10 years, and were of the Murrah, Jafarabadi, Mediterranean, and Murrah × Mediterranean crossbreed breeds. The animals were raised without mineral supplementation. The clinical signs observed in the animals included acromotrichia and depigmentation, with varying degrees and distribution of skin lesions. Histological examination of the epidermis showed interrupted melanin production, mild dermal fibrosis, mild perivascular mononuclear inflammatory infiltrate, and pigmentary incontinence. None of the animals had the genotype for albinism. After 120 days of mineral supplementation based on the use of copper sulfate, the clinical signs of leucoderma regressed. There was no predisposition by breed, sex, or age for the occurrence of the disease. The regression of skin lesions after proper mineral supplementation suggests that copper deficiency may be considered an important factor for the occurrence of leucoderma in buffaloes in the Amazon biome.

An investigative and epidemiological study was carried out for equine herpesvirus type 1 (HVE-1) in 10 outbreaks of neurological disease from different farms in the state of Pará, Brazil. 25 horses were studied: six male and 19 females, aged between one and 13 years. A necropsy of six horses was performed, and the others recovered either with or without treatment (T1-vitamin B1 + dexamentasone; T2-vitamin B1 + flunixim meglumine). Animals that received treatment recovered after eight days. The main clinical signs observed were motor incoordination, progressive paresis, thoracic and/or pelvic limbs abducted after induction of clinical examination, knuckling of the hind fetlocks, sagging and swaying of the hindquarters while standing or walking and paresis. All animals were positive: 88% (22/25) in nested PCR and 72% (18/25) in serum neutralization (including three negatives in serology). Focal brownish areas compatible with hemorrhage were found in the white and gray matter of the spinal cord of two animals. On histological analysis, there were perivasculitis and neutrophilic vasculitis in the gray matter of the spinal cord and brain. Based on the evidence, this work proves the circulation of HVE-1 in the Amazon biome, mainly in the state of Pará, Brazil.

Transcatheter pulmonary valve replacement has become an attractive alternative to surgical approach in patients with dysfunctional right ventricular outflow tract. However, in certain cases, an unfavorable anatomy might complicate optimal valve deployment and stability. Several techniques have been described to reshape the landing zone and allow proper implantation of the transcatheter valve. Among them, the hybrid approach has gained attention as an interesting method for off-pump pulmonary valve replacement in patients with dilated right ventricular outflow tract. But to date, there is no standardized method to resize and reshape the landing zone for the stented valve. Here, we describe a reproducible method based on simple geometric rules to allow adequate remodeling of the main pulmonary artery to the desired dimensions in a single attempt, followed by perventricular implantation of a Venus P-valve.

The alpha (CPA), beta (CPB) and epsilon (ETX) toxins of Clostridium perfringens are responsible for causing diseases that are difficult to eradicate and have lethal potential in production animals. Vaccination of herds is still the best control strategy. Recombinant clostridial vaccines have shown good success at inducing neutralizing antibody titers and appear to be a viable alternative to the conventional production of commercial clostridial toxoids. Research is still needed on the longevity of the humoral immune response induced by recombinant proteins in immunized animals, preferably in target species. The objective of this study was to measure the humoral immune response of cattle immunized with trivalent vaccines containing the recombinant proteins alpha (rCPA), beta (rCPB) and epsilon (rETX) of C. perfringens produced in Escherichia coli at three different concentrations (100, 200, and 400 µg) of each protein for 12 months. The recombinant vaccines containing 200 (RV2) and 400 µg (RV3) yielded statistically similar results at 56 days. They performed better throughout the study period because they induced higher neutralizing antibody titers and were detectable for up to 150 and 180 days, respectively. Regarding industrial-scale production, RV2 would be the most economical and viable formulation as it achieved results similar to RV3 at half the concentration of recombinant proteins in its formulation. However, none of the vaccines tested induced the production of detectable antibody titers on day 365 of the experiment, the time of revaccination typically recommended in vaccination protocols. Thus, reiterating the need for research in the field of vaccinology to achieve greater longevity of the humoral immune response against these clostridial toxins in animals, in addition to the need to discuss the vaccine schedules and protocols adopted in cattle production.

Botulism is a neuroparalytic intoxication, usually fatal, caused by the botulinum toxins (BoNTs). Vaccination is the best-known strategy to prevent this disease in ruminants. Serotypes C and D and their variants CD and DC are the main types responsible for botulism in bovine and buffaloes in Brazil and cattle in Japan and Europe. Brazil has a herd of approximately 1.39 million buffaloes and is the largest producer in the Western world. This study aimed to assess the humoral immune response of buffaloes during the 12-month period after vaccination against BoNT serotypes C and D with a recombinant vaccine in three different concentrations (100, 200, and 400 μg) of non-purified recombinant proteins (Vrec) and also with a bivalent commercial toxoid (Vcom). Vrec400 was the best vaccine among those tested because it induced higher levels of antibodies and maintained higher levels of antibodies for the longest time, while Vrec200 could be considered the most cost-effective vaccine for large-scale production. None of the vaccines were able to promote continuous immunological protection within the timeframe proposed by the current Brazilian vaccination protocol. Further studies should focus on vaccine adjustments to ensure continued humoral protection against botulism., Competing Interests: Declaration of competing interest None., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Propolis is a resinous substance composed of a mixture of different plant parts and molecules secreted by bees. Chemically, it is defined as a complex matrix containing biologically active molecules with antibacterial, antifungal, antiviral, antiparasitic, hepatoprotective, and immunomodulatory activities. It is widely employed in cosmetic formulations and pharmaceutical products and is one of the most widely used natural products. However, the effects and strength of these biological activities depend on the chemical profile and composition of each propolis type. This composition is associated with the diversity of local flora, the place and period of collection, and the genetics of the bees. In this context, the objective of this review was to investigate the biological, chemical, and microbiological properties of propolis. A technological prospection was also performed on patents for products designed to be used in animal health. Our investigation shows that the literature contains diverse studies dedicated to comparing and describing the composition and therapeutic properties of propolis. These studies demonstrate the potential biological use of propolis in veterinary medicine, showing the applications of propolis extracts in different formulations. However, there are a low number of propolis-based veterinary products with a registered patent. Thus, the development of products based on propolis is a promising market to be exploited. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Mutations in the CLCN1 gene are the primary cause of non-dystrophic Hereditary Myotonia in several animal species. However, there are no reports of Hereditary Myotonia in pigs to date. Therefore, the objective of the present study was to characterize the clinical and molecular findings of Hereditary Myotonia in an inbred pedigree. The clinical, electromyographic, histopathological, and molecular findings were evaluated. Clinically affected pigs presented non-dystrophic recessive Hereditary Myotonia. Nucleotide sequence analysis of the entire coding region of the CLCN1 gene revealed the absence of the exons 15 and 16 in myotonic animals. Analysis of the genomic region flanking the deletion unveiled a large intragenic deletion of 4,165 nucleotides. Interestingly, non-related, non-myotonic pigs expressed transcriptional levels of an alternate transcript (i.e., X2) that was identical to the deleted X1 transcript of myotonic pigs. All myotonic pigs and their progenitors were homozygous recessive and heterozygous, respectively, for the 4,165-nucleotide deletion. This is the first study reporting Hereditary Myotonia in pigs and characterizing its clinical and molecular findings. Moreover, to the best of our knowledge, Hereditary Myotonia has never been associated with a genomic deletion in the CLCN1 gene in any other species.

Clostridium perfringens type A is the causative agent of gas gangrene and gastroenteric ("yellow lamb disease") disease in ruminants, with C. perfringens alpha toxin (CPA) being the main virulence factor in the pathogenesis of these illnesses. In the present study, we have developed recombinant Escherichia coli bacteria expressing rCPA and used it to vaccinate rabbits and sheep. Doses of up to 200 μg of rCPA used for inoculation, induced 13.82 IU.mL -1 of neutralizing antitoxin in rabbits, which is three times higher than that recommended by the USDA (4 IU.mL -1 ). In sheep, recombinant bacteria induced antitoxin titers of 4 IU.mL -1 , 56 days after the first dose. rCPA which was expressed, mainly, in inclusion bodies, was not found to influence the immunogenicity of the vaccine. The recombinant Escherichia coli bacterin, produced simply and safely, is capable of affording protection against diseases caused by C. perfringens CPA. The current findings represent a novel production method for CPA vaccines potentially applicable to veterinary medicine., (Copyright © 2019. Published by Elsevier Ltd.)

Ethnopharmacological Relevance: The oleoresin of Brazilian Copaifera reticulata is a traditional remedy used for the treatment of skin and urinary tract infections, respiratory diseases, rheumatism, ulcer and tumours; thus, playing an important role in the primary health care of the indigenous population., Aim: As most previous pharmacological tests used the crude oleoresin and only a few studies so far dealt with enriched fractions or pure chemically defined compounds, the aim of this study was to systematically evaluate the antimicrobial and cytotoxic properties of the Copaifera reticulata oleoresin and to assign traditional uses to specific secondary metabolites., Materials and Methods: The oleoresin, as well as its neutral and acidic fractions were tested for their activity against six cancer cell lines, two clinically relevant bacterial strains, and two dermatophytes. Both fractions were analysed by GC-MS and UHPLC-ELSD, respectively. The antibacterial acidic phase was further fractionated by preparative chromatography to purify and characterize the compounds responsible for the observed pharmacological effect., Results: Whereas no cytotoxic activity was detected, the crude oleoresin and its acidic fraction showed antibacterial activity against gram-positive bacteria Enterococcus faecium (IC 50 values 4.2 and 4.8 µg/mL, respectively) and methicillin-resistant Staphylococcus aureus (MRSA, IC 50 values 5.3 and 7.2 µg/mL, respectively). Purification of the acidic fraction of the C. reticulata oleoresin yielded two dicarboxylic diterpene acids and the four main diterpene acids, comprising three different diterpene scaffolds. Interestingly, the activity was not restricted to a particular diterpene-type but rather depended on the compounds' lipophilicity, with the most active constituent showing IC 50 values of 1.6 (E. faecium) and 2.5 µg/mL (MRSA), respectively. Furthermore, ent-polyalthic acid, the major diterpenoid, was significantly active against dermatophytes with IC 50 values of 6.8 µg/mL (Trichophyton rubrum) and 4.3 µg/mL against (T. mentagrophytes)., Conclusion: The present study proved the antimicrobial effects of the C. reticulata oleoresin and its diterpenoid constituents, confirming its wide use in folk medicine for the treatment of skin and urinary tract infections. The inhibitory activity of copaiba diterpenoids against dermatophytic fungi as well as the gram-positive bacteria E. faecium and MRSA is being reported for the first time, providing potential lead structures for the treatment of these clinically relevant bacterial strains., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Aphids, like most animals, mount a diverse set of defenses against pathogens. For aphids, two of the best studied defenses are symbiont-conferred protection and transgenerational wing induction. Aphids can harbor bacterial symbionts that provide protection against pathogens, parasitoids and predators, as well as against other environmental stressors. In response to signals of danger, aphids also protect not themselves but their offspring by producing more winged than unwinged offspring as a way to ensure that their progeny may be able to escape deteriorating conditions. Such transgenerational wing induction has been studied most commonly as a response to overcrowding of host plants and presence of predators, but recent evidence suggests that pea aphids (Acyrthosiphon pisum) may also begin to produce a greater proportion of winged offspring when infected with fungal pathogens. Here, we explore this phenomenon further by asking how protective symbionts, pathogen dosage and environmental conditions influence this response. Overall, while we find some evidence that protective symbionts can modulate transgenerational wing induction in response to fungal pathogens, we observe that transgenerational wing induction in response to fungal infection is highly variable. That variability cannot be explained entirely by symbiont association, by pathogen load or by environmental stress, leaving the possibility that a complex interplay of genotypic and environmental factors may together influence this trait., Competing Interests: Nicole Gerardo, one of the authors of this manuscript, was a PLOS ONE editor until 2014. This does not alter the authors’adherence to all the PLOS ONE policies on sharing data and materials.

Botulism is a potentially fatal intoxication caused by botulinum neurotoxins (BoNTs) produced mainly by Clostridium botulinum . Vaccination against BoNT serotypes C and D is the main procedure to control cattle botulism. Current vaccines contain formaldehyde-inactivated native BoNTs, which have a time-consuming production process and pose safety risks. The development of non-toxic recombinant vaccines has helped to overcome these limitations. This study aims to evaluate the humoral immune response generated by cattle immunized with non-purified recombinant fragments of BoNTs C and D. Cattle were vaccinated in a two-dose scheme with 100, 200 and 400 µg of each antigen, with serum sampling on days 0, 56, 120, and 180 after vaccination. Animals who received either 200 or 400 μg of both antigens induced titers higher than the minimum required by the Brazilian ministry of Agriculture, Livestock and Food Supply and achieved 100% (8/8) seroconversion rate. Animals vaccinated with commercial toxoid vaccine had only a 75% (6/8) seroconversion rate for both toxins. Animals that received doses containing 400 µg of recombinant protein were the only ones to maintain titers above the required level up until day 120 post-vaccination, and to achieve 100% (8/8) seroconversion for both toxins. In conclusion, 400 µg the recombinant Escherichia coli cell lysates supernatant was demonstrated to be an affordable means of producing an effective and safe botulism vaccine for cattle.

We report, for the first time, the presence of ungulate malaria parasites in South America. We conducted PCR-based surveys of blood samples of multiple deer species and water buffalo from Brazil and detected Plasmodium sequences from pampas deer ( Ozotoceros bezoarticus ) samples. Phylogenic analysis revealed that the obtained sequences are closely related to the Plasmodium odocoilei clade 2 sequence from North American white-tailed deer ( Odocoileus virginianus ). Nucleotide differences suggest that malaria parasites in South American pampas deer and North American P. odocoilei clade 2 branched more recently than the Great American Interchange.

Botulism is a fatal intoxication caused by botulinum neurotoxins (BoNTs), which are mainly produced by Clostridium botulinum and characterized by flaccid paralysis. The BoNTs C and D are the main serotypes responsible for botulism in animals, including buffaloes. Botulism is one of the leading causes of death in adult ruminants in Brazil due to the high mortality rates, even though botulism in buffaloes is poorly reported and does not reflect the real economic impact of this disease in Brazilian herds. Vaccination is reported as the most important prophylactic measure for botulism control, although there are no specific vaccines commercially available for buffaloes in Brazil. This study aimed to evaluate the humoral immune response of buffalo groups vaccinated with three different concentrations of recombinant proteins (100, 200, and 400 µg) against BoNTs serotypes C and D as well as to compare the groups to each other and with a group vaccinated with a bivalent commercial toxoid. The recombinant vaccine with a concentration of 400 μg of proteins induced the highest titers among the tested vaccines and was proven to be the best choice among the formulations evaluated and should be considered as a potential vaccine against botulism in buffalo., Competing Interests: The authors declare no conflict of interest.

The objective of this study was to determine the prevalence of antibodies to Neospora caninum and Toxoplasma gondii among 500 cattle (Bos indicus) and 500 buffaloes (Bubalus bubalis) using the indirect fluorescent antibody test (IFAT) technique. Blood samples from were collected from water buffalo and cattle in 10 municipalities in the northern region of Brazil. The frequency of cattle and water buffaloes seropositive for Neospora caninum in Pará state, Brazil, was 55% and 44%, respectively, and the frequency of cattle and water buffaloes seropositive for Toxoplasma gondii was 52% and 39%, respectively. Seropositivity for both N. caninum and T. gondii was detected in 10.6% of the cattle samples and 14.8% of the buffalo samples. The frequency of cattle positive for N. caninum and T. gondii was significantly (p<0.05) higher than that of buffalo in two and three provinces, respectively. Buffaloes had a lower seroprevalence for N. caninum or T. gondii in all of the provinces studied. These results suggest that both species, when exposed to the same risks for N. caninum and T. gondii infection, have a high serological prevalence. Cattle showed a higher probability of being seropositive when exposed to the same risks for N. caninum and T. gondii. Our study, which included an extensive number of blood samples, provides important epidemiological information pertinent to buffalo production in tropical countries that can be used as a basis for disease-management practices in Latin America., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Propolis is known for its biological properties and its preparations have been continuously investigated in an attempt to solve the problem of their standardization, an issue that limits the use of propolis in food and pharmaceutical industries. The aim of this study was to evaluate in vitro antioxidant, antimicrobial, antiparasitic, and cytotoxic effects of extracts of red, green, and brown propolis from different regions of Brazil, obtained by ethanolic and supercritical extraction methods. We found that propolis extracts obtained by both these methods showed concentration-dependent antioxidant activity. The extracts obtained by ethanolic extraction showed higher antioxidant activity than that shown by the extracts obtained by supercritical extraction. Ethanolic extracts of red propolis exhibited up to 98% of the maximum antioxidant activity at the highest extract concentration. Red propolis extracts obtained by ethanolic and supercritical methods showed the highest levels of antimicrobial activity against several bacteria. Most extracts demonstrated antimicrobial activity against Staphylococcus aureus. None of the extracts analyzed showed activity against Escherichia coli or Candida albicans. An inhibitory effect of all tested ethanolic extracts on the growth of Trypanosoma cruzi Y strain epimastigotes was observed in the first 24 h. However, after 96 h, a persistent inhibitory effect was detected only for red propolis samples. Only ethanolic extracts of red propolis samples R01Et.B2 and R02Et.B2 showed a cytotoxic effect against all four cancer cell lines tested (HL-60, HCT-116, OVCAR-8, and SF-295), indicating that red propolis extracts have great cytotoxic potential. The biological effects of ethanolic extracts of red propolis revealed in the present study suggest that red propolis can be a potential alternative therapeutic treatment against Chagas disease and some types of cancer, although high activity of red propolis in vitro needs to be confirmed by future in vivo investigations.

In 2011, an outbreak of severe vesicular disease occurred in the state of Pará, Amazon region. Besides proliferative or verrucous lesions, cattle showed atypical clinical signs such as diarrhea and leading to death. The animals were submitted to clinical, pathological and molecular diagnosis, and laboratory tests have confirmed the presence of Pseudocowpox virus (PCPV), a Parapoxvirus genus member, and have also found Bovine viral diarrhea virus-1 (BVDV-1), probably causing persistent infection. The results of molecular diagnostics, followed by sequencing data demonstrated the circulation of both viruses (PCPV and BVDV-1) in an area previously affected by another poxvirus, as Vaccinia virus.The cocirculation between PCPV and BVDV-1 indicates a major concern for animal health because the clinical presentation can be a severe disease. This is the first detection of PCPV in the Brazilian Amazon., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: The presence of lymphoma in buffaloes was first reported in India in the 1960s. The disease is similar to Enzootic Bovine Leucosis (EBL) caused by Bovine leukemia virus (BLV) in cattle; however, according to our results and those of other studies, the etiology of these lymphomas in buffalo do not appear to be associated with BLV. The objectives of this study are to describe four cases of the disease in buffaloes belonging to the same herd in the Amazon region of Brazil and to perform a clinical-anatomopathological, immunohistochemical, and etiological study of the lymphomas., Results: Over a period of ten years, four buffaloes were observed presenting progressive weight loss, swelling of peripheral lymph nodes, and nodules in the subcutaneous tissue. Upon necropsy, whitish-colored tumor masses were observed in the form of nodules in the subcutaneous tissue, along with miliary nodules on the serosal surfaces of abdominal and thoracic organs and tumors in lymph nodes and other organs. Neoplastic lymphocyte proliferation was observed through histopathology. An immunohistochemical study revealed that the neoplasias were formed by proliferation of predominantly B lymphocytes. The presence of BLV genome was not detected in the lymphomas when using the real-time PCR technique, nor was it detected through immunohistochemical staining using monoclonal antibodies against two viral proteins. Bovine herpesvirus 6 was not detected in the tumors. However, Bovine immunodeficiency virus (BIV) was detected in samples of lymphoma and in the lymph nodes and kidneys of one of the animals., Conclusions: The occurrence of lymphoma in buffaloes is reported for the first time in Brazil and is characterized by B-cell multicentric lymphoma. The etiology of the disease does not appear to be associated with BLV; however, the detection of BIV in samples of lymphoma from one sick animal deserves further study, considering the oncogenic potential of this virus.

Several farms in the Northeast of Brazil were investigated for Mycobacterium avium subsp. paratuberculosis infection in order to identify the occurrence of paratuberculosis in buffaloes. Samples were obtained from 17 farms, two slaughter houses, and a quarantine area in the Northeast. About 15,000 buffaloes of the Murrah, Mediterranean, and Jafarabadi breed as well as their crossbreeds were evaluated for meat, dairy, and mixed farms with semi-intensive or extensive breeding practices. For diagnostic purposes, postmortem and histopathological examination, including Ziehl-Neelsen test of fecal smears and scraped intestinal mucosa were performed. PCR was applied for fecal samples, mesenteric lymph nodes, and intestines. Six Johne's disease-positive farms, which together with those previously identified, indicate that the disease is spread through the Brazilian Northeast, similar to what occurs in cattle herds in other regions of the country. The increase in prevalence of paratuberculosis is a consequence of introduction of animals from other regions without adequate veterinary assistance and due to the little official attention paid to this initially silent and chronic disease.

Approximately 50% of buffalo herds in Brazil are located in Pará state in northern Brazil. There are several properties where cattle and buffalo live and graze together, and thus, buffalo pathogens may threaten the health of cattle and vice versa. Therefore, knowledge of infectious agents of buffalo is essential for maintaining healthy livestock. Clinical disease caused by Theileria and Babesia parasites in the Asian water buffalo is not common, although these animals may act as reservoir hosts, and the detection of these hemoparasites in buffaloes is as important as it is in cattle. Studies of the infection of buffaloes by hemoparasites in Brazil are scarce. The objective of the present study was to investigate the occurrence of Piroplasmida parasites in Asian water buffaloes in the state of Pará in the Amazon region of Brazil using nested PCR assays and phylogenetic analysis. The 18S rRNA gene and ITS complete region were amplified from DNA extracted from blood samples collected from 308 apparently healthy buffaloes bred on six properties in the state of Pará, Brazil. The prevalence of positive buffalo samples was 4.2% (13/308) for Theileria spp., 3.6% (11/308) for Babesia bovis and 1% (3/308) for Babesia bigemina. Animals infected with Theileria were detected in 50% (3/6) of the assessed properties. Phylogenetic analyses indicated that the Theileria species detected in this study were closely related to Theileria buffeli, Theileria orientalis and Theileria sinensis. To our knowledge, this is the first report of Theileria in Asian water buffaloes in the Americas. The majority of Theileria-positive buffaloes (11/13) belong to a property that has a history of animals presenting lymphoproliferative disease of unknown etiology. Therefore, the present research suggests that this disorder can be associated with Theileria infection in this property. Our results provide new insights on the distribution and biological aspects of hemoparasites transmissible from buffaloes to cattle., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Enzootic bovine leucosis is an infectious disease caused by Bovine leukemia virus (BLV) and is well described in bovines. The majority of infected animals are asymptomatic, one to five percent develop lymphoma and from 30 to 50% present a persistent lymphocytosis. The virus occurs naturally in cattle and experimentally in buffaloes, capybaras and rabbits. The occurrence of lymphoma in buffaloes has been attributed to BLV infection by some authors in India and Venezuela, but not confirmed by other studies and little information on natural BLV infection in buffaloes is available. The aim of this study was to evaluate the occurrence of BLV in a sub-sample of buffalo from Amazon and southeast regions in Brazil. Three hundred and fifteen serum samples were negative using commercial AGID and ELISA (ELISA-gp51) which detect anti-BLV glycoprotein gp51 antibodies. The same samples were also evaluated for antibodies to whole virus through a commercial ELISA (ELISA-BLV) in which 77 (24.44%) were found seropositive and two (0.63%) inconclusive. On the other hand, all animals were negative by PCR to BLV targeted to the env and tax genes. These results suggest that ELISA-BLV produces false positive results in buffalo serum (p<0.001). In addition, one buffalo lymphoma sample was negative in both PCR assays used in this study. BLV was not detected in buffaloes from the Amazon basin and the southeast region of Brazil. Serological tests, like ELISA-BLV, usually used for cattle may produce false-positive results for BLV in buffaloes and direct detection tests such as PCR should be chosen in these surveys. The occurrence of lymphoma in buffalo was not associated with BLV infection in the one case analyzed in this work and the etiology and pathogenesis of this disease should be clarified., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In Brazil, serologic evidence of Orthopoxvirus (OPV) circulation showed positivity around 20% in cattle, humans, monkeys and rodents. Although OPV seropositivity has been described in buffalo herds in southeastern Brazil, no Vaccinia virus (VACV) (member of genus OPV) outbreaks in buffalo herds have been described in this country. This study aimed to investigate the detection of anti-OPV antibodies and to study the OPV genome in Brazilian buffalo herds. Our results demonstrated a high OPV seropositivity in buffalo herds on Marajó Island and molecular data confirmed the circulation of VACV. The geographical isolation conditionmight be a sine qua non condition to explain our results., (Copyright © 2016. Published by Elsevier Ltd.)

Bovine immunodeficiency is a chronic progressive disease caused by a lentivirus that affects cattle and buffaloes. Although the infection has been described in cattle in some countries, including in Brazil, there are only two reports of infection in buffaloes: one in Pakistan and one in Cambodia. The aim of the present study was to survey the occurrence of bovine immunodeficiency virus (BIV) in water buffaloes from the Amazon region, Pará state, Brazil. BIV proviral DNA was surveyed in 607 whole blood samples of water buffaloes from 10 farms located in the state of Pará using semi-nested polymerase chain reaction (PCR) (PCR-SN) to amplify the pol region of the viral genome. Of the 607 samples tested, 27 (4.4 %) were positive for BIV proviral DNA. The amplified fragments were confirmed by sequence analysis after cloning and nucleotide sequencing. The sequence obtained had 99 % similarity to the reference strain (R-29). The present study provides important epidemiological data because BIV was detected for the first time in water buffaloes in Brazil. Further, the results suggest the possibility of the virus being a risk factor for herd health because it may be a potential causal agent of chronic disease and, also may be associated to other infectious diseases.

Anaplasma marginale is the most prevalent pathogen transmitted by ticks in cattle in tropical and subtropical regions of the world. However, the tick species involved in the transmission of A. marginale in buffaloes in Brazil have not been identified. The objective of the present study was to determine the presence of A. marginale in ticks parasitizing water buffaloes. A total of 200 samples of Rhipicephalus microplus, Dermacentor nitens, Amblyomma cajennense, and Amblyomma maculatum were collected and tested by conventional and quantitative PCR for the presence of the msp1a and msp5 genes. In the present study, 35 ticks (17.5%) were positive for A. marginale DNA by qPCR analysis. The positive ticks belonged to four different species: R. microplus (22.2%), A. cajennense (13.8%), A. maculatum (16.0%), and D. nitens (10.0%). Individuals of the three developmental stages (larvae, nymphs, and adults) of R. microplus and A. cajennense were found to be positive for A. marginale, only nymphs and adults of A. maculatum were found to be positive, and finally, only adults of D. nitens were positive for A. marginale. Our results suggest that R. microplus, A. cajennense, A. maculatum, and D. nitens ticks may be involved in the transmission of A. marginale in buffaloes. However, while A. marginale PCR positive ticks were recorded, this does not indicate vector competence; only that the ticks may contain a blood meal from an infected host. Additionally, the results show that the strains of A. marginale from buffaloes and cattle are phylogenetically related., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Tuberculosis is a disease with a great zoonotic potential. It is considered a major obstacle to cattle production and is responsible for severe losses in several production systems. A comparative cervical test (CCT) was performed in 1140 buffaloes from different mesoregions of the state of Pará, Brazil, with the aim of comparing the sensitivity and specificity of CCT with histopathological examination and bacterial culture. Of the animals tested using CCT, 4.65% (53/1140) were positive, 2.98% (34/1140) were inconclusive, and 92.36% (1053/1140) were negative. Among the 168 sacrificed animals, 33 were positive, 18 were inconclusive, and 117 were negative by CCT, and samples from the sacrificed animals were collected for histopathological examination and bacterial culture. A qualitative evaluation of the tuberculin test was performed by comparing the test results with the histopathological and bacteriological results. The latter two tests yielded a prevalence of 4.16%, a sensitivity of 71.43%, and a specificity of 82.61%. Based on these results, we concluded that CCT yielded satisfactory results and can be applied in diagnostic studies in buffaloes. The prevalence rate obtained using three distinct diagnostic methods suggests that Mycobacterium bovis was present in a few animals in the population evaluated.

Background: Transcatheter closure of a moderate to large patent ductus arteriosus (PDA) using conventional techniques is challenging. The Nit-Occlud® PDA-R trial can close a PDA up to 8 mm in diameter. We sought to report procedural and six-month efficacy and safety results of the multicenter Nit-Occlud® PDA-R trial., Methods: From June 2010 to February 2011, 43 patients were enrolled in three centers from Argentina. Median age was 4.5 (range 1.4-18.4 years) years old at catheterization, 70% were females and weight was 17.7 (range 10-67 kg)., Results: PDAs mean diameter was 2.98 ± 1.03 and ranged from 2 to 6.19 mm. About 11.6% were large (≥4 mm), whereas 32.6% were <2.5 mm. Median pulmonary artery mean pressure was 17 mm Hg (range 9-26 mm Hg). The device was implanted successfully in all patients. By echocardiography, trivial residual shunt was observed in 42% at the end of the procedure, in 28% at 24 hr, in 12.1% at one week, and none at three-months. There was one case of embolization (due to undersizing), that was treated successfully with a larger study device. There were no major short- or long-term complications., Conclusions: PDAs ranging from 2 to 6 mm can be effectively and safely closed using the Nit-Occlud® PDA-R device, with good procedural and six-month results. The Nit-Occlud® PDA-R emerges as an optimal alternative for closure of small to moderate PDAs., (© 2015 Wiley Periodicals, Inc.)

This study presents the first description of Bovine herpesvirus 6 (BoHV-6) that was isolated from buffaloes of Amazon region in Brazil. Phylogenetic analysis showed that the BoHV-6 Brazilian strains clustered with the sequence of BoHV-6 from elsewhere available at the GenBank. It was observed in some buffaloes with lymphoproliferative disease in one herd, thus the animals were also tested for Bovine leukemia virus (BLV), which has been associated to lymphoma in bovines. All animals were negative to BLV. These results indicate that BoHV-6 is present in buffaloes in Brazil, but the importance and impact of this infection and its association with any illness is still undefined.

Anaplasma marginale is the most prevalent pathogen of cattle in tropical and subtropical regions of the world and causes the disease bovine anaplasmosis. The importance of water buffalo in the world economy is increasing. In addition, while water buffalo may serve as a reservoir host for A. marginale, the susceptibility of this host for A. marginale cattle strains in Brazil has not been reported. The major surface protein 1 alpha (msp1α) gene has been shown to be a stable genetic marker for identification of A. marginale strains. Herein, we analyzed blood samples from 200 water buffalo and identified the A. marginale strains in an endemic area of Rio de Janeiro, Brazil, where ticks were present and water buffalo and cattle co-mingled. Ticks that were feeding on the study buffalo were collected and identified. The prevalence of A. marginale in water buffalo in this study was low (10%). Sequence analysis of the msp1α gene demonstrated the presence of 8 different A. marginale strains. Two A. marginale strains in the water buffalo, (α-β-β-β-Γ) and (α-β-β-Γ), were similar to those reported in cattle from nearby regions. The results of this study suggested that water buffalo in this region are naturally infected with the same strains of A. marginale found in cattle., (Copyright © 2014 Elsevier B.V. All rights reserved.)

All life cycle stages of the protozoan parasite Trypanosoma cruzi are enveloped by mucin-like glycoproteins which, despite major changes in their polypeptide cores, are extensively and similarly O-glycosylated. O-Glycan biosynthesis is initiated by the addition of αGlcNAc to Thr in a reaction catalyzed by Golgi UDP-GlcNAc:polypeptide O-α-N-acetyl-d-glucosaminyltransferases (ppαGlcNAcTs), which are encoded by TcOGNT1 and TcOGNT2. We now directly show that TcOGNT2 is associated with the Golgi apparatus of the epimastigote stage and is markedly downregulated in both differentiated metacyclic trypomastigotes (MCTs) and cell culture-derived trypomastigotes (TCTs). The significance of downregulation was examined by forced continued expression of TcOGNT2, which resulted in a substantial increase of TcOGNT2 protein levels but only modestly increased ppαGlcNAcT activity in extracts and altered cell surface glycosylation in TCTs. Constitutive TcOGNT2 overexpression had no discernible effect on proliferating epimastigotes but negatively affected production of both types of trypomastigotes. MCTs differentiated from epimastigotes at a low frequency, though they were apparently normal based on morphological and biochemical criteria. However, these MCTs exhibited an impaired ability to produce amastigotes and TCTs in cell culture monolayers, most likely due to a reduced infection frequency. Remarkably, inhibition of MCT production did not depend on TcOGNT2 catalytic activity, whereas TCT production was inhibited only by active TcOGNT2. These findings indicate that TcOGNT2 downregulation is important for proper differentiation of MCTs and functioning of TCTs and that TcOGNT2 regulates these functions by using both catalytic and noncatalytic mechanisms., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

The rickettsia Anaplasma marginale is the etiologic agent of bovine anaplasmosis, an important tick-borne disease affecting cattle in tropical and subtropical regions of the world. In endemic regions, the genetic diversity of this pathogen is usually related to the high prevalence of the disease in cattle. The major surface protein 1 alpha (MSP1a) has been used as a marker to characterize the genetic diversity and for geographical identification of A. marginale strains. The present study reports the characterization of A. marginale MSP1a diversity in water buffaloes. Blood samples were collected from 200 water buffaloes on Marajó Island, Brazil where the largest buffalo herd is located in the Western hemisphere. Fifteen buffaloes (7.5%) were positive for A. marginale msp1α by PCR. Four different strains of A. marginale with MSP1a tandem repeat structures (4-63-27), (162-63-27), (78-24-24-25-31) and (τ-10-10-15) were found, being (4-63-27) the most common. MSP1a tandem repeats composition in buffalos and phylogenetic analysis using msp1α gene showed that the A. marginale strains identified in buffaloes are closely related to A. marginale strains from cattle. The results demonstrated low genetic diversity of A. marginale associated with low bacterial prevalence in buffaloes and suggested that buffaloes may be reservoirs of this pathogen for cattle living in the same area. The results also suggested that mechanical transmission and not biological transmission by ticks might be playing the major role for pathogen circulation among water buffaloes in Marajó Island, Brazil., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

Bovine anaplasmosis, caused by Anaplasma marginale, occurs in tropical and subtropical regions throughout the world and is a major constraint on cattle production in many countries. Approximately 60% of the buffalo herds in South America are located in northern Brazil. However, compared with the research on cattle, research on buffaloes has been neglected. Therefore, the present study was conducted to investigate the distribution of A. marginale in water buffaloes in northern Brazil. A total of 500 buffalo blood samples was randomly collected from 16 provinces and was analyzed using both nPCR assay and ELISA techniques. The percentage of animals that were seropositive for A. marginale according to ELISA was 49% (245/500). The main risk factors associated with seroprevalence were the region (p=0.021; OR=1.2) and the reproductive status (p=0.0001; OR=1.6). Anaplasma marginale DNA was detected in 5.4% (27/500) of the sampled buffaloes. Our data provide information about the incidence of A. marginale infection in water buffaloes and may guide future programs aimed at controlling the disease in the northern region of Brazil. Although these water buffaloes are exposed to A. marginale, a low rate of A. marginale PCR-positive animals was found, which could be explained by the habitat in which the sampled animals live because they exhibited a low rate of attached ticks on their skin., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

The prevalence of and possible risk factors for tuberculosis were studied in water buffalo from Pará, Brazil. In this study, 3,917 pregnant and nonpregnant female Murrah and Mediterranean buffaloes were studied; 2,089 originated from Marajó Island, and 1,108 were from the mainland. The comparative cervical tuberculin test was used as a diagnostic test for tuberculosis in these animals. The prevalence of positive buffaloes was 3.5 % (100/2,809) on Marajó Island and 7.2 % (80/1,108) on the mainland. The municipalities with the highest tuberculosis prevalence rates in animals were Ipixuna do Pará (10.1 %), Marapanim (9.8 %), Chaves (9.4 %), Paragominas (8.6 %), and Cachoeira do Arari (6.7 %). The tuberculosis prevalence was not significantly different between the Murrah (4.3 %) and Mediterranean (4.8 %) breeds or between pregnant (5 %) and nonpregnant (4.3 %) buffaloes. Tuberculosis was detected in water buffaloes from Pará, Brazil; the mainland buffalo exhibited the highest tuberculosis prevalence. These results indicate that this disease is dangerous to public health and buffalo farming in Pará.

To evaluate the prevalence and possible risk factors for brucellosis caused by Brucella abortus in water buffaloes in the state of Pará, Brazil, 3,917 female buffalo serum samples from pregnant and non-pregnant animals were examined: 2,809 from Marajó Island and 1,108 from the mainland. The buffered acidified plate antigen (BAPA) screening test positively diagnosed 4.8% (188/3,917) of the animals with brucellosis, and the 2-mercaptoethanol (2-ME) confirmatory test affirmed 95.7% (180/188) of the results. The brucellosis prevalence was 4.17 times greater in mainland animals than on Marajó Island, with the highest prevalence in Tailândia (11.30%) and Paragominas (12.38%). Brucellosis seroprevalence was significantly influenced (p < 0.05) by reproductive status, with pregnant females being most vulnerable. These results demonstrate that brucellosis infection is active in the Brazilian region containing the largest buffalo population and that this disease poses a threat to public health and buffalo production in Pará.

Although the largest buffalo herd in the occident is in the north region of Brazil, few studies have been conducted to assess the prevalence of selected parasitic diseases in buffalo herd. The present study was therefore conducted to investigate the epidemiological of Toxoplasma gondii, Neospora caninum, Anaplasma marginale, Babesia bigemina, and Babesia bovis in water buffaloes in the north region of Brazil. A total of 4796 buffalo blood samples were randomly collected from five provinces and simultaneously analyzed by the IFAT and ELISA. The serological prevalence of T. gondii and N. caninum was 41.3% and 55.5% in ELISA and 35.7% and 48.8% in IFAT, respectively. The overall prevalence of A. marginale, B. bovis, and B. bigemina was 63%, 25%, and 21% by ELISA and 50.0%, 22.5%, and 18.8% by IFAT, respectively. This study shows valuable information regarding the serological survey of selected bovine pathogens in water buffaloes in the north region of Brazil which will likely be very beneficial for the management and control programs of this disease.

In 2010, vaccinia virus caused an outbreak of bovine vaccinia that affected dairy cattle and rural workers in Pará State, Brazil. Genetic analyses identified the virus as distinct from BeAn58058 vaccinia virus (identified in 1960s) and from smallpox vaccine virus strains. These findings suggest spread of autochthonous group 1 vaccinia virus in this region.