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Introduction: Bovine viral diarrhea virus (BVDV), a positive-sense single-stranded RNA virus, causes significant economic losses in the cattle industry. Current diagnostic methods for BVDV exhibit variable sensitivity and specificity, underscoring the need for more rapid and accurate detection approaches. Here, we developed a novel competitive ELISA (cELISA) to detect antibodies against the BVDV E2 protein. Methods and results: We generated three monoclonal antibodies (mAbs)—3E6, 2D5, and 5B9—by immunizing mice with purified BVDV E2 protein expressed in Expi293F cells. Among these, mAb 3E6 displayed superior competitive binding abilities to the E2 protein, enabling effective differentiation between BVDV positive and negative sera. Remarkably, mAb 3E6 exhibited pan-genotypic recognition of various BVDV strains, including BVDV-1a, -1b, -1c, -1m, -1p, -1v, and -2a, while showing no cross-reactivity with the classical swine fever virus (CSFV). Computational modeling using AlphaFold 3 identified domain B of the E2 protein as the primary binding site for mAb 3E6. Building upon these findings, we established a cELISA employing mAb 3E6 and recombinant E2 protein. Receiver-operating characteristic (ROC) analysis revealed outstanding diagnostic performance, achieving a sensitivity of 99.26% and specificity of 98.99%. Further tests confirmed the cELISA's specificity for detecting BVDV-specific antibodies, with no cross-reactivity with antisera from animals infected or immunized against BCoV, BHV-1, BRV, AKAV, LSDV, BLV, and CSFV. Consistency was observed between results from the BVDV E2 cELISA and traditional virus neutralization test (VNT), demonstrating high sensitivity for monitoring antibody dynamics. In performance evaluations, the established cELISA exhibited high concordance with VNT in assessing 160 vaccinated sera and 190 clinical samples. Discussion: The BVDV E2 cELISA, utilizing mAb 3E6 to target domain B of the BVDV E2 protein, represents a reliable and effective serological diagnostic tool for the detection of antibodies against both BVDV-1 and BVDV-2. This methodology holds significant promise for applications in clinical diagnosis and the evaluation of vaccine efficacy. [ABSTRACT FROM AUTHOR]

There are inconclusive claims in the scientific literature that the species Trema micranthum, widely distributed throughout the Brazilian territory, may produce phytocannabinoids, potentially serving as an alternative to Cannabis sativa. In this study, we conducted a comprehensive investigation to assess the presence of phytocannabinoids in two Trema micranthum samples collected in the Midwest region of Brazil. In trying to detect cannabinoids in T. micranthum, a recommended cannabis screening test was employed, the Fast Blue BB Salt (FBBBS) colorimetric assay, followed by thin-layer chromatography (TLC) and instrumental techniques: high-performance liquid chromatography coupled to diode array detector (HPLC-DAD) and gas chromatography coupled to mass spectrometry (GC-MS). When employed without chloroform extraction, the FBBBS reagent yielded positive results for extracts from all parts of T. micranthum (leaves, branches, fruits, and inflorescences). However, these initial positive results from the FBBBS test, suggesting the presence of cannabinoids, were not corroborated by FBBBS followed by chloroform extraction, TLC, or the instrumental techniques used in this study. These additional outcomes suggest that the positive FBBBS test results were likely due to the presence of other phenolic compounds rather than phytocannabinoids. For example, the presence of vitexin-like compounds in T. micranthum extracts might explain the positive FBBBS test results. Therefore, new assertions that T. micranthum produces cannabinoids will require the support of more selective experiments to avoid false-positive claims based on less selective screening tests. [ABSTRACT FROM AUTHOR]

Brain abscesses in ruminants often arise from primary infection foci, leading to an unfavorable prognosis for affected animals. This highlights the need for comprehensive studies on brain abscesses across different ruminant species. We retrospectively investigated medical records of epidemiological, clinical, neuroimaging, anatomopathological, and bacteriological findings in six ruminants (three goats, two cows, and one sheep) diagnosed with brain abscesses. All animals studied were female. Apathy (50%), compulsive walking (33%), decreased facial sensitivity (33%), head pressing (33%), seizures (33%), semicomatous mental status (33%), strabismus (33%), unilateral blindness (33%), and circling (33%) represented the most common neurologic signs. Leukocytosis and neutrophilia were the main findings in the hematological evaluation. Cerebrospinal fluid (CSF) analysis revealed predominant hyperproteinorrachia and pleocytosis. In three cases, computed tomography or magnetic resonance imaging were used, enabling the identification of typical abscess lesions, which were subsequently confirmed during postmortem examination. Microbiological culture of the abscess samples and/or CSF revealed bacterial coinfections in most cases. Advanced imaging examinations, combined with CSF analysis, can aid in diagnosis, although confirmation typically relies on postmortem evaluation and isolation of the causative agent. This study contributes to clinicopathological aspects, neuroimages, and bacteriological diagnosis of brain abscesses in domestic ruminants. [ABSTRACT FROM AUTHOR]

Opportunistic bacteria and fungi are commonly reported causes of bovine abortion in a small percentage of fetal losses of infectious etiology in cattle. The objective of this study was to characterize the pathological and etiological findings in fetuses aborted due to secondary bacterial and fungal infections submitted for postmortem examination between 2004 and 2019 in the State of Rio Grande do Sul, Brazil. Nineteen cases of bacterial etiology and five cases of fungal etiology were assessed. In cases of bacterial etiology, gross changes were uncommon and two different microscopic patterns were observed: (1) primary bronchopneumonia with occasional dissemination in cases of Staphylococcus sp., Streptococcus sp., and Mannheimia haemolytica infections; and (2) systemic disease with sepsis in cases of Escherichia coli and Listeria sp. infections. Aspergillus sp. was the main fungal agent identified, and cases of mycotic abortion were characterized by placentitis, dermatitis, and pneumonia. Fetal membranes were available for examination in less than half of the submissions (11/24), and placental lesions were observed in all cases. This study reaffirms the importance of postmortem examinations in the determination of causes of fetal loss in cattle and highlights pathological findings commonly observed in fetuses aborted due to sporadic bacterial and fungal agents., (© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Viral neurologic diseases are common in cattle, although most non-suppurative meningoencephalitis (NSM) remains etiologically unknown. We compared the epidemiological, clinical, and pathological data among 79 cases of rabies, 12 cases of NSM of unknown etiology (NSM-UE), and 8 cases of herpetic meningoencephalitis previously diagnosed in cattle in Southern Brazil. Neurological clinical signs were similar among rabies and NSM-UE and different in cattle with herpetic meningoencephalitis. Only two herpetic meningoencephalitis cases had gross lesions in the central nervous system, characterized by malacia and hemorrhage. Histologically, all three groups had mild to severe multifocal infiltrates of lymphocytes, plasma cells, and macrophages/microglial cells in the Virchow-Robin space, neuropil, and leptomeninges, and gliosis. Other findings included malacia and eosinophilic intracytoplasmic inclusion in rabies, and malacia and intranuclear amphophilic inclusion in herpetic meningoencephalitis. By immunohistochemistry, the predominant inflammatory cells in all cases were T lymphocytes, followed by macrophages/microglial cells, B lymphocytes, and astrocytes. The T lymphocyte count showed statistically significant differences between the diseases. Our results revealed few differences between the groups. Although the etiological agent involved has not been identified in cases of NSM-UE, the characteristics observed in this study showed similarity with viral diseases., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Bovine viral diarrhea virus (BVDV) is the causative agent of bovine viral diarrhea (BVD), which results in significant economic losses in the global cattle industry. Fortunately, various diagnostic methods available for BVDV have been established. They include etiological methods, such as virus isolation (VI); serological methods, such as enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay (IFA), and immunohistochemistry (IHC); molecular methods, such as reverse transcription-polymerase chain reaction (RT-PCR), real-time PCR, digital droplet PCR (ddPCR), loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), and CRISPR-Cas system; and biosensors. This review summarizes the current diagnostic methods for BVDV, discussing their advantages and disadvantages, and proposes future perspectives for the diagnosis of BVDV, with the intention of providing valuable guidance for effective diagnosis and control of BVD disease. [ABSTRACT FROM AUTHOR]

Simple Summary: Buffalo farming has become economically important in Brazil given the highly rustic nature of the species and the better nutritional characteristics of buffalo meat and milk. However, the sector has been impacted by the occurrence of different diseases in buffaloes that have been poorly described in the international literature to date. This study reports and describes the clinically observed neurological signs and the macroscopic and microscopic lesions of bacterial meningitis in adult buffaloes raised in the Amazon biome. These lesions are associated with fractures of the base of the horn and exposure of the frontal sinus. 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. [ABSTRACT FROM AUTHOR]

Simple Summary: Mortality within the nursery pig herd constitutes a pivotal factor contributing to economic losses and concurrently serves as a valuable indicator of production efficiency. The correct diagnosis is an important tool for guiding management corrections, improvements in animal performance, and decision making in herds. Thus, this study aimed to establish the main causes of death in nursery pigs from different nurseries through necropsies. Eighteen nurseries were analyzed (a total of 120,243 housed piglets), and 557 necropsies were performed. Conclusive diagnosis totaled 93.2% (519/557). Bacterial and viral diseases constituted the majority of conclusive diagnoses (72.9%) (378/519), followed by undefined causes at 14.4% (75/519) and non-infectious diseases at 12.7% (66/519). There was an increase in mortality rate in individual nurseries associated mainly with bacterial disease outbreaks. Therefore, infectious diseases constituted the largest portion of the diagnoses, offering a great opportunity for improving production rates in herds. In this sense, the aspects highlighted of the main causes of death can guide accurate diagnosis and specific decision making in nurseries. Necropsies can reveal herd problems or comorbidities that can lead to management corrections, improvements in animal performance, and better decision making. Furthermore, the pattern and causes of mortality might differ when different systems are evaluated. The present study was conducted to establish the main causes of death in nursery pigs from different systems in Brazil, as well as the clinical, microbiological, and pathological aspects of these mortalities. Eighteen nurseries were analyzed (a total of 120,243 housed piglets), and 557 necropsies were performed. Streptococcus suis infection was the most prevalent cause of death (21.2%), followed by bacterial polyserositis (16.7%), chronic atrophic enteritis (13.5%), salmonellosis (8.8%), pneumonia (8.6%), and colibacillosis (6.1%). The increase in mortality rate in individual nurseries and, consequently, in the diagnoses was commonly associated with disease outbreaks. Infectious diseases constituted the largest portion of the diagnoses, making a great opportunity for improving production rates in herds. Moreover, the extensive range of observed diagnoses highlights the importance of conducting preliminary diagnostic investigations based on necropsy to determine the causes of death. This approach allows for the direction of complementary tests, which can diagnose agents with greater specificity. As a result, this allows for the implementation of more effective prevention and control strategies. [ABSTRACT FROM AUTHOR]

Toxoplasmosis is an important zoonotic disease caused by the parasite Toxoplasma gondii and is especially fatal for neotropical primates. In Brazil, the Ministry of Health is responsible for national epizootic surveillance, but some diseases are still neglected. Here, we present an integrated investigation of an outbreak that occurred during the first year of the COVID-19 pandemic among eleven neotropical primates housed at a primatology center in Brazil. After presenting non-specific clinical signs, all animals died within four days. A wide range of pathogens were evaluated, and we successfully identified T. gondii as the causative agent within four days after necropsies. The liver was the most affected organ, presenting hemorrhage and hepatocellular necrosis. Tachyzoites and bradyzoite cysts were observed in histological examinations and immunohistochemistry in different organs; in addition, parasitic DNA was detected through PCR in blood samples from all specimens evaluated. A high prevalence of Escherichia coli was also observed, indicating sepsis. This case highlights some of the obstacles faced by the current Brazilian surveillance system. A diagnosis was obtained through the integrated action of researchers since investigation for toxoplasmosis is currently absent in national guidelines. An interdisciplinary investigation could be a possible model for future epizootic investigations in animals. [ABSTRACT FROM AUTHOR]

Astroviruses (AstVs) have been detected in a wide range of animal species, including mammals and birds. Recently, a novel AstV associated with neurological symptoms has been detected in the brains of some mammals. Raccoon dog AstV has been reported recently in China. However, there have been no reports in South Korea. Therefore, the present study aimed to detect and genetically characterize AstVs in the intestine and brain tissues of 133 wild raccoon dogs collected in Korea between 2017 and 2019. Of the seven raccoon dogs, AstVs were detected in six intestinal tissues and four brain tissues. Analysis of the capsid protein amino acid sequences of raccoon dog AstVs detected in Korea revealed a high similarity to canine AstVs, suggesting possible interspecies transmission between raccoon dogs and dogs. Phylogenetic and capsid protein amino acid sequence analysis of raccoon dog AstVs detected in the brain the 17-148B strain belonging to the HMO clade and exhibiting conserved sequences found in neurotropic AstVs (NT-AstVs), indicating their potential as NT-AstVs. However, the pathogenicity and transmission routes of the raccoon dog AstV detected in Korea have not yet been elucidated, so further research and continued surveillance for AstV in wild raccoon dogs are needed. [ABSTRACT FROM AUTHOR]

The photolysis of colchicine under ultraviolet and visible light irradiation was studied by ultraviolet (UV) scanning and HPLC-MS. The photoproduct was proposed and the cytotoxicity change before and after irradiation was investigated. Results showed that both ultraviolet and visible light irradiation could effectively degrade colchicine into deacetamido-lumicolchicine. The process conformed to first-order kinetics, in which a high degradation rate (K = 0.5862 h− 1) was observed when colchicine was dissolved in ethanol and irradiated by UV light. Cell viability and cell cycle studies proved that a photolysis treatment of colchicine could weaken the cytotoxicity effectively. Colchicine inhibited the division of BRL 3 A cells in G2/M phase with an IC50 value of 0.48 µg/mL, while the toxic effect could be reduced significantly with IC50 2.1 µg/mL when colchicine was exposed to UV irradiation. Results are beneficial to the toxicity elimination of colchicine in the processing of daylily flower in food industry, and can also provide photochemistry reference for colchicine-related studies. [ABSTRACT FROM AUTHOR]

Background: The European bison (Bison bonasus) is a near threatened species and requires health monitoring. The aim of the present study was to determine the prevalence of antibodies to pathogens known to cause respiratory and digestive illness in ruminants. Results: In the studied 328 European bison, the highest seroprevalence was observed for Bovine herpesvirus-1 (BoHV-1) (50.27%), Bovine Coronavirus (BCoV) (26.36%), and Bluetongue Virus (BTV) (12.83%). For Mycoplasma bovis strains and Bovine Viral Diarrhea Virus (BVDV), positive results were rare. Interestingly, a higher prevalence of BTV antibodies was noted in the northeastern populations and older animals. Conclusions: Our findings indicate that the Polish European bison population appears to have considerable contact with BoHV-1; however, this does not appear to be of great significance, as clinical symptoms and post-mortem lesions are rarely noted in Polish European bison population. The high seroprevalence of BTV in the north-east of Poland is an ongoing trend, also noted in previous studies. It is possible that European bison may perpetuate the virus in this region. This is the first report of antibodies for BCoV in European bison. [ABSTRACT FROM AUTHOR]

The present study aimed to test intestinal scrapings of the Northern Goshawk (Accipiter gentilis) and the Eurasian Sparrowhawk (Accipiter nisus) from Lithuania for S. calchasi and other Sarcocystis species characterised by bird–bird life cycles. The protozoan parasite Sarcocystis calchasi can cause respiratory and neurological diseases in a variety of birds; however, the distribution of this parasite is not well-examined. Sarcocystis species were identified with nested PCR and sequencing of the partial ITS1 region. Sporocysts and/or sporulated oocysts of Sarcocystis spp. were observed in 16 (100%) Northern Goshawks and 9 (56.3%) Eurasian Sparrowhawks. Four species, S. columbae, S. halieti, S. turdusi, and S. wobeseri, were confirmed in the Eurasian Sparrowhawk. Apart from the latter four species, S. calchasi, S. cornixi, S. kutkienae, and S. lari were established in the Northern Goshawk. A higher prevalence of Sarcocystis spp. and species richness in Northern Goshawks is associated with the differences in the diet of two examined Accipiter species. This study is the first report of S. calchasi in Lithuania. Furthermore, the genetically distinct species Sarcocystis spp. 23LTAcc, which is most closely related to S. calchasi, was found in three Northern Goshawks. [ABSTRACT FROM AUTHOR]

Simple Summary: This paper reports a case of suppurative meningoencephalitis by Pseudomonas aeruginosa from the direct extension of chronic otitis in a Gir cow. The direct extension of otitis media/interna results in secondary pyogranulomatous intracranial lesions occasionally. Clinical signs included forebrain, cerebellum, cranial nerve VII, VIII, and XII impairment. Pure cultures of P. aeruginosa were obtained and identified in the suppurative central nervous system (CNS) lesions, meninges, and inner ear samples. The paucity of case reports of middle or inner ear infections evolving to suppurative processes in the CNS of adult cattle suggests underreporting or lack of diagnosis. This paper aims to report clinical, laboratory and pathological features in a case of suppurative meningoencephalitis by P. aeruginosa from the direct extension of chronic otitis in a Gir cow. The cow was recumbent during physical examination, and neurological examination revealed depression, absence of left eyelid and auricular motor reflex, and hypotonic tongue. Hematology revealed hemoconcentration, leukocytosis by neutrophilia, and hyperfibrinogenemia. Cerebrospinal fluid was slightly turbid, and presented polymorphonuclear pleocytosis, and hyperproteinorrachia. Grossly, the skull floor showed a purulent green–yellow exudate that drained from the left inner ear to the cisterna magna. There was diffuse congestion of the telencephalon, and meninges showed severe hyperemia, moderate thickening, and opacity with the deposition of fibrinosuppurative material ventrally, extending to the cerebellum and brainstem. The left cerebellar hemisphere showed an approximately 1.5 cm in diameter liquefaction area surrounded by a hemorrhagic halo. Histologically, cerebellar, mesencephalic, thalamic, and brain base meninges were intensely thickened and showed severe suppurative inflammation and fibrin deposition. Small multifocal suppurative areas were observed in the cerebellum and brainstem, characterized by a necrotic core, a number of neutrophils, and Gram-negative intralesional bacillary myriads. Pure cultures of P. aeruginosa were obtained and identified in the suppurative CNS lesions, meninges, and inner ear samples. This report highlights an uncommon clinical evolution of secondary P. aeruginosa suppurative meningoencephalitis, probably triggered by recurrent parasitic otitis in an adult Gir cow. Veterinarians, practitioners, and farmers must be aware of the risk of CNS infections after unresolved media and inner otitis, especially in cattle breeds more prone to developing parasitic otitis, such as the Gir and Indubrasil breeds. [ABSTRACT FROM AUTHOR]

The work aims to study the economical losses of the condemnation of bovine and buffalo carcasses, in order to estimate the losses in animals slaughtered in Santarém-Pará, Brazil, between 2016 and 2018, with data obtained from the Municipal Department of Agriculture and Fisheries. Sex, age, origin, total number of animals slaughtered and causes of condemnation of carcasses were considered. All analyzes were performed in RStudio version 1.1.463. In this study, 71,277 bovine carcasses and 2,016 buffalo carcasses were inspected, of which 300 bovine and 71 buffalo were condemned. The highest prevalence of causes of condemnation in cattle was recorded for brucellosis (0.0020%) and tuberculosis (0.0019%). In buffaloes, tuberculosis (0.0307%) peritonitis (0,0019%) were the main causes of condemnations. Economical losses were more evident in females, for both species. The projection of economical losses related to the condemnation of carcasses showed a sharp growth for the next three years, if the average growth remains constant. The biggest projected loss was for bovine females, with an accumulated projection of $ 5,451.44. The smallest estimated loss was for buffalo males, projected at more than thirty-two thousand reais. The most important causes of condemnation report the diseases brucellosis and tuberculosis, as the ones with the greatest impact. In the buffalo species this was even more accentuated, even though the number of buffaloes slaughtered is more than 35 times smaller than the number of cattle. [ABSTRACT FROM AUTHOR]

Background: Central nervous system (CNS) infections in cattle are a major cause of economic loss and mortality. Machine learning (ML) techniques are gaining widespread application in solving predictive tasks in both human and veterinary medicine. Objectives: Our primary aim was to develop and compare ML models that could predict the likelihood of a CNS disorder of infectious or inflammatory origin in neurologically‐impaired cattle. Our secondary aim was to create a user‐friendly web application based on the ML model for the diagnosis of infection and inflammation of the CNS. Animals: Ninety‐eight cattle with CNS infection and 86 with CNS disorders of other origin. Methods: Retrospective observational study. Six different ML methods (logistic regression [LR]; support vector machine [SVM]; random forest [RF]; multilayer perceptron [MLP]; K‐nearest neighbors [KNN]; gradient boosting [GB]) were compared for their ability to predict whether an infectious or inflammatory disease was present based on demographics, neurological examination findings, and cerebrospinal fluid (CSF) analysis. Results: All 6 methods had high prediction accuracy (≥80%). The accuracy of the LR model was significantly higher (0.843 ± 0.005; receiver operating characteristic [ROC] curve 0.907±0.005) than the other models and was selected for implementation in a web application. Conclusion and Clinical Importance: Our findings support the use of ML algorithms as promising tools for veterinarians to improve diagnosis. The open‐access web application may aid clinicians in achieving correct diagnosis of infectious and inflammatory neurological disorders in livestock, with the added benefit of promoting appropriate use of antimicrobials. [ABSTRACT FROM AUTHOR]

Pestiviruses are globally distributed and cause substantial economic losses to the cattle industry. In Brazil, the country with the world's largest cattle population, pestivirus infections are well described in some regions, such as in the south, where a high frequency of BVDV-2 is described and contrasts with the high prevalence of HoBi-like pestivirus (HoBiPeV) in the northeast. However, there is a lack of information about pestiviruses in the Amazon Region, in northern Brazil, with a cattle population estimated at 55.7 million head, which has a significant impact on the international livestock market. Therefore, this study investigated the seroprevalence and genetic variability of ruminant pestiviruses in 944 bovine serum samples from four states in northern Brazil: Pará (PA), Amapá (AP), Roraima (RR), and Amazonas (AM). Our results showed that 45.4% of the samples were seropositive (19.8% for BVDV-1, 14.1% for BVDV-2, and 20.9% for HoBiPeV). All samples were tested by RT–qPCR, and three were positive and classified as HoBiPeV in a phylogenetic analysis. These serological and molecular results contrast with those from other regions of the world, suggesting that the northern Brazilian states have a high prevalence of all bovine pestiviruses including HoBiPeV. [ABSTRACT FROM AUTHOR]

Neutrophils are innate immune cells that play crucial roles in response to extracellular pathogens, including bacteria and fungi. Pythium insidiosum (P insidiosum) is a fungus-like pathogen that causes "pythiosis" in mammals. This study investigated in vitro function of human neutrophils against P. insidiosum. We demonstrated the killing mechanism of neutrophils when incubated with P. insidiosum zoospores (infective stage), such as phagocytosis and neutrophil extracellular traps (NETs). Healthy human neutrophils significantly reduced six strains of live zoospores isolated from different sources compared to the condition without neutrophils (p < 0.001), observed by colony count and trypan blue staining. As our results showed the killing ability of neutrophils, we further investigated the neutrophil killing mechanism when incubating with zoospores. Our study found that only two strains of heat-killed zoospores significantly induced phagocytosis (p < 0.01). Co-culture of heat-killed zoospores and neutrophils demonstrated NET formation, which was detected by immunofluorescence staining using DAPI, anti-myeloperoxidase, and anti-neutrophil elastase and quantitated under the fluorescence microscope. In addition, the level of cell-free DNA released from neutrophils (as a marker of NET production) after incubation with zoospores showed significantly increased levels when compared with unstimulated neutrophils (p < 0.001). Our findings demonstrate that neutrophils revealed the NET formation in response to P. insidiosum zoospores. This study is the first observation of the neutrophil mechanism against P. insidiosum, which could provide a better understanding of some parts of the innate immune response during pythiosis. [ABSTRACT FROM AUTHOR]

Pythiosis, a life-threatening infectious condition caused by Pythium insidiosum, has been increasingly reported in humans and animals worldwide. Antifungal drugs usually fail to control the pathogen. The surgical removal of an infected organ is the treatment of choice. Many affected patients die due to advanced infection. A timely and accurate diagnosis could lead to a better prognosis in pythiosis patients and save their lives. Although a standard culture method is available in microbiological laboratories, it is time-consuming, laborious, and insensitive for P. insidiosum identification. Immunological assays have been developed to improve the diagnosis of pythiosis. However, immunological methods are commercially unavailable and primarily detect anti-P. insidiosum antibodies, which constitute indirect evidence of pythiosis, making it challenging to differentiate a past from a recent infection. Moreover, such immunological tests cannot diagnose patients with a local infection, such as in the eye. Nucleic acid-based tests (NATs) are efficient for the direct and rapid detection of P. insidiosum DNA in trace-amount or culture-negative specimens. The reagents and equipment required for NATs are usually available in molecular diagnostic laboratories. Herein, we provide a systematic review to comprehensively present the principal and clinical usages, advantages, and limitations of such NATs in the detection of P. insidiosum. Various NATs have been established to detect P. insidiosum, which can be classified into amplification-based (i.e., PCR assays, isothermal tests, and next-generation sequencing methods) and non-amplification-based (i.e., DNA hybridization) techniques. This concise review on NATs constitutes an up-to-date reference with which healthcare professionals can learn about and decide upon which detection method is suitable for their respective laboratory environments. [ABSTRACT FROM AUTHOR]

Rabies is a viral, lethal, and zoonotic disease with worldwide distribution, primarily transmitted through bites from infected dogs and bats. The Biosafety Laboratory (LSE-LANASEVE) of the Animal Health Service of Costa Rica (SENASA) serves as the national reference laboratory for diagnosing rabies in humans and animals. Since regional laboratories lack the equipment for the direct fluorescent antibody test (FAT), we evaluated the rapid immunochromatographic diagnostic test (RIDT) from BioNote, employing FAT as a reference, to improve rabies diagnosis. We analyzed 193 brain tissue samples between 2014 and 2019. Out of these, 174 came from species that RIDT has been validated for: bovines (162), dogs (10), and raccoons (2). The rest were from unvalidated species, including horses (7), humans (1), and others. Among the 174 validated samples, 26 bovine samples were positive for both RIDT and FAT. Reviewing all 193 samples, 28 were positive and 165 negative using both methods. Two horse samples presented inconsistencies, being positive on FAT but negative on RIDT; these were subsequently verified as false negatives by RT-PCR. RIDT exhibited a sensitivity of 94 % (CI95, 83.9-102.3), specificity of 100 %, positive predictive value (PPV) of 100 %, and negative predictive value (NPV) of 99 % (CI95, 97.1-100.5). RIDT has demonstrated reliability in quickly diagnosing rabies for validated species. We advise its application in SENASA’s regional laboratories for those particular species. If there’s uncertainty, samples should be sent to LSE-LANASEVE for FAT or RT-PCR confirmation. [ABSTRACT FROM AUTHOR]

Bovine viral diarrhea virus (BVDV) belongs to the family Flaviviridae genus pestivirus. The viral genome is a single-stranded, positive-sense RNA that encodes four structural proteins (i.e., C, Erns, E1, and E2) and eight non-structural proteins (NSPs) (i.e., Npro, p7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B). Cattle infected with BVDV exhibit a number of different clinical signs including diarrhea, abortion, and other reproductive disorders which have a serious impact on the cattle industry worldwide. Research on BVDV mainly focuses on its structural protein, however, progress in understanding the functions of the NSPs of BVDV has also been made in recent decades. The knowledge gained on the BVDV non-structural proteins is helpful to more fully understand the viral replication process and the molecular mechanism of viral persistent infection. This review focuses on the functions of BVDV NSPs and provides references for the identification of BVDV, the diagnosis and prevention of Bovine viral diarrhea mucosal disease (BVD-MD), and the development of vaccines. [ABSTRACT FROM AUTHOR]

Sarcocystis spp. are apicomplexan cyst-forming parasites that can infect numerous vertebrates, including birds. Sarcosporidiosis infection was investigated in three muscles (breast, right and left thigh muscle) and one organ (heart) of four Razorbill auks (Alca torda) stranded between November and December 2022 on the shores of the Mediterranean Sea in Nabeul and Bizerte governorates, Northern Tunisia. Two of the four tested A. torda were PCR positive for 18S rRNA Sarcocystis spp. gene. Among the examined 16 muscles/organs, only one breast and one right thigh were Sarcocystis spp. PCR-positive (12.5% ± 8.3, 2/16). Our results showed a relatively high molecular prevalence of Sarcocystis spp. in Razorbill auks (A. torda). Sarcocystis spp. sequence described in the present study (GenBank number: OR516818) showed 99.56-100% identity to Sarcocystis falcatula. In conclusion, our results confirmed the infection of Razorbill auks (A. torda) by S. falcatula. Further research is needed on different migratory seabirds' species in order to identify other Sarcocystis species., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Pythiosis is a disease caused by the oomycete Pythium insidiosum, mainly reported in equines, dogs and humans and directly transmitted through contaminant zoospores in aquatic environments. We report the first outbreak of equine pythiosis in five equines. Wound samples were submitted for diagnostic testing including mycological culture and nested PCR. Treatment approaches consisted of conventional and alternative therapies. Microbiological analyses were performed using water samples from the riverbanks close to where the animals had grazed. All animals were positive for P. insidiosum cultures, and two animals responded successfully to alternative therapy (ozone therapy). After culture and molecular analysis of environmental samples, the presence of P. insidiosum in one section of the Tietê River was confirmed through a 99% sequence identity. Phylogenetic analyses using the cytochrome oxidase II gene showed that the animal isolates clustered in clade I and the environmental isolates clustered in clade III. Although the environmental and wound isolates belonged to different genetic clades, we concluded that the Tietê River is an important source of infection by P. insidiosum and that research concerning environmental isolation of P. insidiosum from rivers and lakes should be strongly facilitated in Brazil. [ABSTRACT FROM AUTHOR]

Background: Botulism is a disease caused by the ingestion of neurotoxin produced by Clostridium botulinum, characterized by flaccid paralysis, which can lead to high mortality. They have seven types of neurotoxins (A, B, C, D, E, F, and G) and, in birds, most cases are attributed to type C. They are considered sources of botulinum toxins where the decomposition of organic matter occurs, like stagnant water and rotting food. The main feature of the disease in birds is ascending symmetric flaccid paralysis. The present study aims to describe an outbreak of type C botulism in backyard poultry in the state of Santa Catarina, Southern Brazil. Case: A visit was made to the property with 160 backyard poultry with a history of high mortality in the municipality of Agrolândia, Santa Catarina. Clinical signs were characterized by paralysis of the pelvic limbs, neck and pendular wings, which progressed to death within 48 h. There was a mortality rate of 37.5% (60/160) between March and May 2019. These birds were kept in an overcrowded environment, with different species (chickens, ducks, teals, and turkeys) fed irregularly. The water supplied was provided from kitchen exhaust, accumulating in puddles on the floor that contained organic matter residues such as animal feces, food waste and bone fragments. The disposal of the carcasses of birds that died was in the same enclosure, buried superficially, facilitating the access of other birds to dig them up and consume them. Necropsy was performed on 2 chickens and one duck, no macroscopic or histopathological lesions were observed. Blood, liver, and gastrointestinal content samples were sent for research and identification of botulinum toxin through the serum neutralization test in mice. The presence of type C botulinum toxin was confirmed in the liver chicken of one sampled animals. Discussion: The identification of type C botulism toxin enabled the characterization of the outbreak, which is the toxin most associated with episodes of botulism in birds. It is not always possible to identify the origin of the infection, as intoxication can occur by ingestion of water contaminated with organic waste, however, in this outbreak, as sources of poisoning in birds, it was specified and occurred due to the ingestion of water with organic matter that was stagnant in the floor of the enclosure, and also by ingesting contaminated carcasses present in the area. In subsistence farming, botulism outbreaks are reported with greater frequency in the Northeast and Southeast of Brazil generally, cases in which sanitary conditions and incorrect carcass disposal favor the occurrence of the disease. As noted in the present study, high mortality is a common feature of botulism. The evaluated signs and developed evolution were similar to previous studies, which ranged from 14 to 72 h. The absence of macroscopic and histopathological changes is commonly reported in cases of botulism in domestic animals, since botulinum toxin only causes functional changes, with no tissue damage. The association of clinical signs, epidemiology, post mortem evaluation and detection of type C botulinum toxin concludes the diagnosis of botulism. Avian influenza and Newcastle disease are important diseases that have neurological conditions and high mortality that should be distinguished from botulism. This report confirms the need to associate history, clinical signs, absence of lesions with laboratory research for the diagnosis of botulism in birds. In addition, it reinforces the importance of disclosing basic management measures to prevent the occurrence of outbreaks such as presented in this report. [ABSTRACT FROM AUTHOR]

Five psittacine birds, one eastern rosella (Platycercus eximius), one rose-ringed parakeet (Psittacula krameri), two eclectus parrot (Eclectus roratus), and one princess parrot (Polytelis alexandrae), all housed in a commercial aviary from La Plata, Buenos Aires, Argentina, suddenly died after a short period of dyspnea. The most significant histopathological findings for all specimens were interstitial exudative pneumonia, with marked congestion and hemorrhage, septa thickening, and massive perivascular lymphoplasmacytic infiltration. Structures compatible with protozoal schizonts were observed in the capillary lumen. No bacterial development was obtained and the real-time PCR for Chlamydia spp. and several psittacine viruses were negative. All the samples resulted negative on the specific PCR for T. gondii. Sarcocystis spp. PCR was positive in the lung and/or liver samples from all birds. The samples showed a restriction pattern of S. neurona and of S. falcatula-like by PCR–RFLP using JNB25-JD396 and JNB33-JNB54 primers, respectively. Sequences obtained from Sarcocystis sp. 18S rRNA and COI gene from 4 birds showed a high identity among them. The 18S rRNA fragment and complete gene sequences obtained showed the highest similarity with S. falcatula and S. speeri sequences but also with S. neurona SN5 isolate sequence. Likewise, COI sequences have 99.89–100% similarity with S. falcatula and S. speeri sequences. Based on all biological and molecular information recorded, we conclude that the etiological agent was S. falcatula-like, close related with the species shed by opossums in South America. [ABSTRACT FROM AUTHOR]

Minipigs are commonly utilized in dermal toxicology studies, necessitating documentation of background findings, including neoplastic lesions. We describe a case of a Yucatan minipig with a squamous cell carcinoma at the mucocutaneous junction of the lip. The neoplasm appeared grossly as a slightly raised area of skin with ulceration. Histologically, there were nests and trabeculae of neoplastic squamous epithelial cells undergoing keratinization. Multifocally, rafts of these cells were within lymphatic vessels. Squamous cell carcinoma has not previously been reported in Yucatan or other laboratory minipigs, however, has been reported uncommonly in nonlaboratory pot-bellied pigs. Although squamous cell carcinoma has been associated with ultraviolet exposure or papillomavirus in various species, this was unable to be confirmed in this case. [ABSTRACT FROM AUTHOR]

Pythiosis is a disease caused by the oomycete Pythium insidiosum, mainly reported in equines, dogs and humans and directly transmitted through contaminant zoospores in aquatic environments. We report the first outbreak of equine pythiosis in five equines. Wound samples were submitted for diagnostic testing including mycological culture and nested PCR. Treatment approaches consisted of conventional and alternative therapies. Microbiological analyses were performed using water samples from the riverbanks close to where the animals had grazed. All animals were positive for P. insidiosum cultures, and two animals responded successfully to alternative therapy (ozone therapy). After culture and molecular analysis of environmental samples, the presence of P. insidiosum in one section of the Tietê River was confirmed through a 99% sequence identity. Phylogenetic analyses using the cytochrome oxidase II gene showed that the animal isolates clustered in clade I and the environmental isolates clustered in clade III. Although the environmental and wound isolates belonged to different genetic clades, we concluded that the Tietê River is an important source of infection by P. insidiosum and that research concerning environmental isolation of P. insidiosum from rivers and lakes should be strongly facilitated in Brazil., (© 2021 Wiley-VCH GmbH.)

This review aims to update the knowledge of the epidemiology of Bovine viral diarrhoea virus (BVDV) in Indonesia and Southeast Asia and provide a perspective on the control options for BVDV in the Indonesian cattle population in the future. Studies on BVDV in Indonesia, since its first report in that country, and the updated beef and dairy cattle industries are reviewed. In ten of 34 provinces, BVDV is endemic. The subgenotypes of BVDV-1a and BVDV-1c are predominant in Indonesian cattle. However, BVDV is currently not a priority disease to control in Indonesia. Cattle imports from Australia appear to be potentially the most significant source of transmission of BVDV into native cattle, but the control of BVDV conducted in the local quarantine facilities is currently not achieving the aim of controlling BVDV; thus, complementary measures are needed. With the small-scale nature of the vast majority of cattle breeding in the country, the control of BVDV in provinces in which cattle breeding is economically essential may need to be organised by regional and provincial governments. Gaps in our knowledge of BVDV are identified in this review, and strategies for the control of BVDV in Indonesia are discussed. [ABSTRACT FROM AUTHOR]