ABSTRACT
The fact that SARS-CoV-2 has reportedly infected companion, livestock and wildlife animals may constitute a significant risk for virus reservoirs, ground for emerging variants and potential for novel reverse zoonosis. Hence, SARS-CoV-2 surveillance in animal species is crucial to prevent emerging variants which may spread to humans. The present study aimed to develop a simple, high-throughput and ultrafast magnetic bead immunoassay to detect anti-SARS-CoV-2 nucleocapsid and spike reactive IgG antibodies in dog and cat serum samples. The assays were validated using serum from eleven dogs and cats which had SARS-CoV-2 infections confirmed by real-time RT-PCR. The negative cohort consisted of pre-pandemic dog and cat samples. The assays performed at 73-82% sensitivity and 97.5-98% specificity for dogs and 71% sensitivity and 92-94% specificity for cats. The lower assay specificity for cats is explained by the fact that cat pre-pandemic sera showed high levels of cross-reactive with SARS-CoV-2 Nucleocapsid and Spike, supporting that these animals have been exposed to other coronavirus sharing structural similarities with SARS-CoV-2. These assays described in this work are now being used for SARS-CoV-2 surveillance and research purposes.
ABSTRACT
SARS-CoV-2 has caused 775 outbreaks in 29 animal species across 36 countries, including dogs, cats, ferrets, minks, non-human primates, white-tailed deer, and lions. Although transmission from owners to dogs has been extensively described, no study to date has also compared sheltered, foster home and owner dogs and associated risk factors. This study aimed to identify SARS-CoV-2 infection and anti-SARS-CoV-2 antibodies from sheltered, fostered, and owned dogs, associated with environmental and management risk factors. Serum samples and swabs were collected from each dog, and an epidemiological questionnaire was completed by the shelter manager, foster care, and owner. A total of 111 dogs, including 222 oropharyngeal and rectal swabs, tested negative by RT-qPCR. Overall, 18/89 (20.22%) dogs presented IgG antibodies against the N protein of SARS-CoV-2 by magnetic ELISA, while none showed a reaction to the Spike protein. SARS-CoV-2 antibodies showed an age-related association, with 4.16 chance of positivity in adult dogs when compared with young ones. High population density among dogs and humans, coupled with repeated COVID-19 exposure, emerged as potential risk factors in canine virus epidemiology. Dogs exhibited higher seropositivity rates in these contexts. Thus, we propose expanded seroepidemiological and molecular studies across species and scenarios, including shelter dogs.
Subject(s)
COVID-19 , Cat Diseases , Deer , Dog Diseases , Lions , Dogs , Animals , Cats , COVID-19/epidemiology , COVID-19/veterinary , SARS-CoV-2 , Seroepidemiologic Studies , Ferrets , Antibodies, Viral , Mink , Dog Diseases/epidemiologyABSTRACT
Caseous lymphadenitis (CL) in sheep is a chronic contagious disease caused by Corynebacterium pseudotuberculosis, commonly characterized by abscess formation in peripheral lymph nodes and disseminated infections. Nonetheless, other microorganisms, including with zoonotic relevance, can be isolated from CL-resembling lymph nodes. Currently, mycobacteria have been reported in visceral granulomatous lesions in small ruminants, a fact that poses a public health issue, particularly in slaughtered sheep intended for human consumption. Cytology using fine needle aspiration and microbiological culturing are suitable tests for routine diagnostic, whereas present drawbacks and molecular methods have been confirmatory. Data about the occurrence of mycobacteria in both lymph nodes with aspect of CL and apparently healthy visceral nodes of sheep slaughtered for human consumption are scarce. In this study, 197 visceral lymph nodes of sheep showed lymphadenitis and 202 healthy visceral lymph nodes of slaughtered sheep intended for human consumption were submitted to conventional bacteriological diagnosis, mycobacteria culturing, and cytological evaluation. Compatible Corynebacterium isolates were subjected to multiplex PCR targeting 16S rRNA, rpoB, and pld genes to detect C. pseudotuberculosis. Based on microbiological identification, C. pseudotuberculosis (86/197; 43.7%), streptococci γ-hemolytic (17/197; 8.6%), and Trueperella pyogenes (12/197; 6.1%) were prevalent in lymph nodes with abscesses, as opposed to staphylococci (53/202; 26.2%) in apparently healthy lymph nodes. No mycobacteria were isolated. Cytology identified 49.2% (97/197) Gram-positive pleomorphic organisms (coryneform aspect). Multiplex PCR confirmed genetic material of C. pseudotuberculosis in 74.4% (64/86) of the samples with C. pseudotuberculosis isolation and 66% (64/97) samples with cytological coryneform aspect (κ = 86.78%; 95% CI = 79.87-93.68%). These findings emphasize the prevalence of C. pseudotuberculosis in abscess formation among peripheral lymph nodes of sheep. Other bacteria were also identified in lymph nodes sampled that resembling C. pseudotuberculosis-induced infections that may difficult the diagnosis. Multiplex PCR revealed a valuable assay to detect C. pseudotuberculosis, in addition to routine methods applied to CL-diagnosis. No mycobacteria were identified in lymph nodes sampled, with and without apparent lesions. Nonetheless, due to public health impacts, this pathogen should be considered as a differential diagnosis of C. pseudotuberculosis-induced infections during inspection procedures of slaughtered sheep intended for human consumption.
Subject(s)
Bacteria/genetics , Coinfection/veterinary , Corynebacterium pseudotuberculosis/genetics , Lymph Nodes/cytology , Lymph Nodes/microbiology , Lymphadenitis/microbiology , Lymphadenitis/veterinary , Mycobacterium/genetics , Abattoirs , Animals , Bacteria/classification , Brazil/epidemiology , Coinfection/microbiology , Cross-Sectional Studies , Farms , Female , Male , Prevalence , RNA, Ribosomal, 16S/genetics , Random Allocation , Sheep/microbiology , Sheep Diseases/epidemiology , Sheep Diseases/microbiologyABSTRACT
In the enteric nervous system (ENS), nitrergic neurons produce and use nitric oxide (NO) as an inhibitory motor neurotransmitter in response to parasitic infections, including those caused by Toxoplasma gondii. However, damage to the host caused by NO has been reported by various authors, and the role of NO in protection or cytotoxicity continues to be extensively studied. In this study, nitrergic neurons were investigated in the myenteric plexus of the jejunum and the distal colon of rats infected with 500 oocysts of the M7741 strain of T. gondii. Ten rats were randomly assigned into a control group (CG) and infected group (IG; received 500 sporulated oocysts of T. gondii orally). After 24h, the rats were euthanized, and samples of the jejunum and distal colon were obtained and processed for NADPH-diaphorase histochemical analysis. Quantitative and morphometric analysis of the nitrergic neurons in whole mounts containing the myenteric plexus was performed. There was a numeric reduction of nitrergic neurons per mm2 in both jejunum and distal colon. The remaining nitrergic neurons suffered atrophy in the areas of the cell body and nucleus, which resulted in a decrease in cytoplasm. Thus, we conclude that an avirulent strain of T. gondii in a short time causes neuroplastic changes in the small and large intestine of rats.
Subject(s)
Nitrergic Neurons/parasitology , Toxoplasma/pathogenicity , Toxoplasmosis/pathology , Animals , Intestines/innervation , Intestines/parasitology , Myenteric Plexus/parasitology , Myenteric Plexus/pathology , Nitrergic Neurons/pathology , Random Allocation , Rats , Rats, Wistar , Toxoplasma/physiology , VirulenceABSTRACT
Since 1950, the human infection by parasites of the genus Toxocara, characterized by migration and permanence of larvae of the parasite in the liver, lungs, eyes, and brain, have been reported. The route of infection is the ingestion of embryonated eggs present in soil and contaminated vegetables, however there are reports of human illness associated with ingestion of viscera of paratenic hosts, including birds. This work aimed to detect the presence of larvae in the tissues and antibodies in the serum of poultry sold in street markets of Feira de Santana, Bahia, Brazil. Over a period of ten months, 100 birds of four street markets in the city were acquired, being examined organs and muscle tissue for the presence of larvae and the serum for the presence of anti-Toxocara spp. antibodies by indirect enzyme-linked immunosorbent assay (ELISA). Helminth larvae were not found in the examination of pepsin digested tissues, but 80 (89.9%) of 89 bird sera examined presented anti-Toxocara spp. IgY antibodies. These results may indicate that birds are coming from contaminated environments, where man and other animals can also be infected.
ABSTRACT
Toxocariasis is a geohelminth zoonosis with worldwide distribution, mainly transmitted through the ingestion of embryonated eggs of nematodes of the Toxocara genus. The disease can also be transmitted to humans as a result of eating raw or undercooked meat of paratenic hosts, such as chickens. Here, we standardized an enzyme-linked immunosorbent assay (ELISA) for evaluating experimentally the kinetic and avidity index (AI) of IgY in broiler chickens infected with different doses of Toxocara canis eggs (G1:100; G2: 1000; and G3: 5000; n = 12 per group). The test showed 91.7% sensitivity (CI 95%: 77.5-98.3) and 100% specificity (CI 95%: 92.6-100), and highest efficiency (97.0%) at 60 days post infection. Infection was characterized by the presence of high avidity antibodies in the chronic phase. Our results support that the ELISA can be a highly useful tool for the detection of anti-Toxocara antibodies in chickens.
Subject(s)
Antibodies, Helminth/immunology , Enzyme-Linked Immunosorbent Assay , Immunoglobulins/immunology , Toxocara/immunology , Toxocariasis/immunology , Animals , Antibodies, Helminth/analysis , Antibody Affinity , Chickens , Dogs , Enzyme-Linked Immunosorbent Assay/methods , Enzyme-Linked Immunosorbent Assay/standards , Female , Immunoglobulins/analysis , Kinetics , Mice , ROC Curve , Sensitivity and SpecificityABSTRACT
Intestinal epithelial secretion is coordinated by the submucosal plexus (SMP). Chemical mediators from SMP regulate the immunobiological response and direct actions against infectious agents. Toxoplasma gondii is a worldwide parasite that causes toxoplasmosis. This study aimed to determine the effects of chronic infection with T. gondii on the morphometry of the mucosa and the submucosal enteric neurons in the proximal colon of rats. Male adult rats were distributed into a control group (n = 10) and an infected group (n = 10). Infected rats received orally 500 oocysts of T. gondii (ME-49). After 36 days, the rats were euthanized and samples of the proximal colon were processed for histology to evaluate mucosal thickness in sections. Whole mounts were stained with methylene blue and subjected to immunohistochemistry to detect vasoactive intestinal polypeptide. The total number of submucosal neurons decreased by 16.20%. Vasoactive intestinal polypeptide-immunoreactive neurons increased by 26.95%. Intraepithelial lymphocytes increased by 62.86% and sulfomucin-producing goblet cells decreased by 22.87%. Crypt depth was greater by 43.02%. It was concluded that chronic infection with T. gondii induced death and hypertrophy in the remaining submucosal enteric neurons and damage to the colonic mucosa of rats.
Subject(s)
Colon/pathology , Neurons/pathology , Toxoplasmosis, Animal/pathology , Animals , Antibodies, Protozoan/blood , Azure Stains , Cats , Cell Death , Chronic Disease , Colon/innervation , Coloring Agents , Gastrointestinal Agents , Goblet Cells/pathology , Immunoglobulin G/blood , Intestinal Mucosa/cytology , Intestinal Mucosa/innervation , Intestinal Mucosa/pathology , Lymphocytes/immunology , Lymphocytes/pathology , Male , Mice , Myenteric Plexus/cytology , Random Allocation , Rats , Rats, Wistar , Submucous Plexus/cytology , Toxoplasma/immunology , Toxoplasma/pathogenicity , Vasoactive Intestinal PeptideABSTRACT
AIM: To assess the effects of ME-49 Toxoplasma gondii (T. gondii) strain infection on the myenteric plexus and external muscle of the jejunum in rats. METHODS: Thirty rats were distributed into two groups: the control group (CG) (n = 15) received 1 mL of saline solution orally, and the infected group (IG) (n = 15) inoculated with 1 mL of saline solution containing 500 oocysts of M-49 T. gondii strain orally. After 36 d of infection, the rats were euthanized. Infection with T. gondii was confirmed by blood samples collected from all rats at the beginning and end of the experiment. The jejunum of five animals was removed and submitted to routine histological processing (paraffin) for analysis of external muscle thickness. The remaining jejunum from the others animals was used to analyze the general population and the NADH-diaphorase, VIPergic and nitrergic subpopulations of myenteric neurons; and the enteric glial cells (S100-IR). RESULTS: Serological analysis showed that animals from the IG were infected with the parasite. Hypertrophy affecting jejunal muscle thickness was observed in the IG rats (77.02 ± 42.71) in relation to the CG (51.40 ± 12.34), P < 0.05. In addition, 31.2% of the total number of myenteric neurons died (CG: 39839.3 ± 5362.3; IG: 26766.6 ± 2177.6; P < 0.05); hyperplasia of nitrergic myenteric neurons was observed (CG: 7959.0 ± 1290.4; IG: 10893.0 ± 1156.3; P < 0.05); general hypertrophy of the cell body in the remaining myenteric neurons was noted [CG: 232.5 (187.2-286.0); IG: 248.2 (204.4-293.0); P < 0.05]; hypertrophy of the smallest varicosities containing VIP neurotransmitter was seen (CG: 0.46 ± 0.10; IG: 0.80 ± 0.16; P < 0.05) and a reduction of 25.3% in enteric glia cells (CG: 12.64 ± 1.27; IG: 10.09 ± 2.10; P < 0.05) was observed in the infected rats. CONCLUSION: It was concluded that infection with oocysts of ME-49 T. gondii strain caused quantitative and plastic alterations in the myenteric plexus of the jejunum in rats.
Subject(s)
Jejunum/innervation , Muscle, Smooth/innervation , Myenteric Plexus/parasitology , Neuronal Plasticity , Toxoplasma/pathogenicity , Toxoplasmosis/parasitology , Animals , Biomarkers/metabolism , Dihydrolipoamide Dehydrogenase/metabolism , Disease Models, Animal , Male , Myenteric Plexus/metabolism , Myenteric Plexus/physiopathology , Neuroglia/metabolism , Neuroglia/parasitology , Nitrergic Neurons/metabolism , Nitrergic Neurons/parasitology , Rats, Wistar , Time Factors , Toxoplasmosis/physiopathology , Vasoactive Intestinal Peptide/metabolismABSTRACT
O objetivo desse trabalho foi fazer a descrição morfológica e a identificação de Nematoda encontrados em uma estação de tratamento de esgoto de Feira de Santana, BA, Brasil. Foram coletadas 56 amostras de 500 mL em todos os pontos das etapas aeróbias do tratamento. As amostras foram processadas para estudo em microscopia óptica de campo claro (MO) e microscopia eletrônica de varredura (MEV). Foram encontradas larvas L1e L4, classificadas de acordo com o tamanho. As larvas L1 apresentaram as seguintes medidas de comprimento Md = 255,0μm; diâmetro Md = 15,0μm; vestíbulo bucal Md = 10,0μm; esôfago Md = 82,5μm; e cauda Md = 26,2μm. As larvas L4 apresentaram as seguintes medidas de comprimento Md = 740,0μm; diâmetro Md = 42,0μm; vestíbulo bucal Md = 22,5μm; esôfago Md = 200,0μm; e cauda Md = 47,5μm. Fêmeas adultas também foram encontradas, caracterizando-se por apresentam esôfago rabditoide, maturação sexual com presença de ovos na tuba uterina em diferentes fases de formação embrionária. Alguns ovos foram vistos larvados, sugerindo um padrão de larviparidade. Os vermes foram considerados de vida livre e classificadas como da Ordem Rhabditida, Família Strongyloidae e Gênero Strongyloides...
The objective of this work was to morphologically describe and identify Nematoda found in a sewage treatment plant in Feira de Santana, BA, Brazil. A total of 56 samples with 500-mL each were collected at all points in the aerobic treatment stages. The samples were processed for bright field light microscopy (LM) and scanning electron microscopy (SEM). L1 and L4 larvae were found, classified according to size. L1 larvae presented the following measurements: Md=255.0-μm length; Md=15.0-μm diameter; Md=10.0-μm oral vestibule; Md=82.5-μm esophagus; and Md=26.2 μm tail. L4 larvae presented the following measurements: Md=740.0-μm length; Md=42.0-μm diameter; Md=22.5-μm oral vestibule; Md=200.0-μm esophagus; and Md=47.5-μm tail. Adult females have also been found, characterized by rhabditoid esophagus, sexual maturation with presence of eggs in the uterine tube at different stages of embryo formation. Some eggs were seen at larvae stage, suggesting a pattern of larvae parity. The worms were considered as free-living and classified in the Rhabditida order, Strongyloidae Family and Strongyloides genus...
El objetivo de ese trabajo fue realizar la descripción morfológica y la identificación de Nematodo encontrados en una estación de tratamiento de alcantarilla de Feira de Santana, BAHIA, Brasil. Fueron recogidas 56 muestras de 500 mL en todos los puntos de las etapas aerobias del tratamiento. Las muestras fueron procesadas para estudio en microscopía óptica de campo claro (MO) y microscopia electrónica de barrido (MEB). Se encontraron larvas L1 y L4, clasificados según el tamaño. Las larvas L1 presentaron las siguientes medidas de longitud Md=255.0μm; diámetro Md=15.0μm; vestíbulo oral Md=10.0 µm; esófago Md=82,5μm; y cola Md=26.2μm. Las larvas L4 presentaron las siguientes medidas de longitud Md=740.0μm; diámetro Md=42.0μm; vestíbulo oral Md=22,5μm; esófago Md=200.0μm; y cola Md=47.5μm. También se han encontrado hembras adultas, caracterizándose por presentar esófago rabditoide, maduración sexual con presencia de huevos en las trompas uterina en diferentes etapas de la formación del embrión. Algunos huevos fueron vistos larvados, sugiriendo un estándar de larviparidad. Los gusanos fueron considerados de vida libre y clasificados como de Orden Rhabditida, Familia Strongyloidae y género Strongyloides...
Subject(s)
Animals , Helminths/anatomy & histology , Helminths/growth & development , Helminths/physiology , Primary Treatment Plants/analysisABSTRACT
O consumo de verduras in natura pode ser um meio de transmissão de vários parasitos. Este estudo teve como objetivo a avaliação parasitológica de amostras de verduras comercializadas em supermercados e feiras-livres no município de Umuarama PR. Um total de 48 amostras de alface, 40 de couve, 40 de almeirão e 20 de rúcula foram coletadas, com um total de 20 (13,5%) de positivos para Toxocara (5,4%), Ascaris (5,4%), Hymenolepis nana (1,4%), Entamoeba coli (0,7%) e Taenia (0,7%)...
The consumption of fresh vegetables can be a means of transmission of several parasites. This study aims to assess the presence of enteroparasites in samples of vegetables sold in supermarkets and street markets in the city of Umuarama-PR, Brazil. A total of 48 samples of lettuce, 40 samples of cabbage, 40 of curly endive and 20 of rucola were collected, with 20 (13.5%) being positive, for Toxocara (5.4%), for Ascaris (5.4%), for Hymenolepis nana (1.4%), for Entamoeba coli (0.7%) and for Taenia (0.7%)...
El consumo de verduras in natura puede ser un medio de transmisión de varios parásitos. Este estudio ha tenido como objetivo la evaluación parasitológica de muestras de verduras comercializadas en supermercados y ferias libres en el municipio de Umuarama-PR. Un total de 48 muestras de lechuga, 40 de col, 40 de achicoria y 20 de rúcala, han sido recogidas, con un total de 20 (13,5%) de positivos para Toxocara (5,4%), Ascaris (5,4%), Hymenolepis nana (1,4%), Entamoeba coli (0,7%) y Taenia (0,7%)...
Subject(s)
Animals , Intestinal Diseases, Parasitic/diagnosis , Helminthiasis/diagnosis , Helminthiasis/prevention & control , Vegetables/parasitologyABSTRACT
Mice experimentally infected with a pathogenic strain of Leptospira interrogans serovar Canicola produced false negative results (prozone effect) in a microscopic agglutination test (MAT). This prozone effect occurred in several serum samples collected at different post-infection times, but it was more prominent in samples collected from seven-42 days post-infection and for 1:50 and 1:100 sample dilutions. This phenomenon was correlated with increased antibody titres in the early post-infection phase. While prozone effects are often observed in serological agglutination assays for the diagnosis of animal brucellosis and human syphilis, they are not widely reported in leptospirosis MATs.
Subject(s)
Leptospira interrogans serovar canicola/immunology , Leptospirosis/diagnosis , Agglutination Tests , Animals , Antibodies, Bacterial/blood , Antibodies, Bacterial/immunology , Enzyme-Linked Immunosorbent Assay , Female , Humans , Leptospirosis/microbiology , MiceABSTRACT
The aim of this study was to evaluate the effects of chronic infection of Toxoplasma gondii (with genotype I and genotype III strains) on the population density and morphometry of caecal myenteric neurons in rats. Fifteen, 60-day-old, male Wistar rats (Rattus norvegicus) were used. The animals were assigned into three groups: Control Group (CG), Experimental Group 1 (EG1) and Experimental Group 2 (EG2). EG1 animals received 10(5) tachyzoites of the genotype I (BTU IV) T. gondii strain orally, and the EG2 animals received 10(5) tachyzoites of the genotype III (BTU II) strain orally. Thirty days after inoculation, caecal whole-mount preparations were stained by Giemsa technique. The caecal preparations were then analysed by assessing the population density and morphometry of myenteric neurons in specific regions of the caecum: mesenteric apical (MA), antimesenteric apical (AA), antimesenteric basal (AB) and next to caecal ampulla (NA). Myenteric neurons from the AA region were more clustered in EG1 animals (P<0.05). The EG1 animals presented a 16.8% reduction in the area of the nucleus, whereas the EG2 animals showed 18.4% increase (P<0.05). There was a more marked reduction in the cytoplasm of the animals in EG1 (↓23.2%) compared to EG2 (↓6.2%). There was 35.8% neuronal atrophy in the AB region and 16.8% in the region NA of the EG1 animals (P<0.05). In conclusion, different strains of T. gondii cause morphometric changes in caecal myenteric neurons of rats. Only the genotype I strain was able to cause neuronal density changes.
Subject(s)
Cecum/innervation , Neurons/pathology , Toxoplasma/physiology , Toxoplasmosis, Animal/pathology , Animals , Atrophy , Cecum/parasitology , Cecum/pathology , Cell Count , Cell Nucleus/pathology , Cytoplasm/pathology , Male , Myenteric Plexus/cytology , Neurons/parasitology , Random Allocation , Rats , Rats, WistarABSTRACT
Several studies have demonstrated that the myenteric plexus experiences quantitative and morphometric changes in rats inoculated orally with Toxoplasma gondii. This paper aims to verify if these alterations are also seen when the same animals are inoculated intraperitoneally with the parasite. In order to do that, six Wistar rats (Rattus norvegicus) 60 days of age were infected intraperitoneally with 10(6) tachyzoites of a genotype I T. gondii strain (BTU IV). After 60 days, the animals were anaesthetised and underwent laparotomy. All organs from the small and large intestines were removed, measured, dissected and underwent whole-mount Giemsa technique to stain the neurons in the myenteric plexus. A quantitative and morphometric analysis of these cells was made, and it showed that the parasite causes the death of myenteric neurons in the jejunum and morphometric alterations in these cells throughout the intestine. However, the cellular response of myenteric neurons to T. gondii is heterogeneous compared the different organs from the gut.
Subject(s)
Intestines/innervation , Myenteric Plexus/pathology , Neurons/pathology , Toxoplasma/physiology , Toxoplasmosis, Animal/pathology , Animals , Antibodies, Protozoan/blood , Dogs , Immunoglobulin G/blood , Intestines/parasitology , Male , Neurons/parasitology , Random Allocation , Rats , Rats, Wistar , Toxoplasma/immunology , Toxoplasmosis, Animal/parasitologyABSTRACT
The aim of this paper was to genetically characterize Toxoplasma gondii isolates from free range chickens in regions of Brazilian territory in the state of Mato Grosso do Sul (MS) where T. gondii strains have never been studied. In total, T. gondii isolates from 22 free range chickens were included in this study. Fifty chickens from Eldorado, thirty from Rio Verde and ten from Aquidauana were sampled between January and April 2007. In relation to the genetic diversity of T. gondii isolates from chickens in MS, the magnitude of the diversity in the isolates sampled in this study was comparable to the overall diversity in a composite data set. These 22 isolates in MS revealed 11 genotypes, whereas the 321 isolates ever genotyped in Brazil have revealed 95 genotypes. The values of Simpson's Diversity Index for the whole population of T. gondii isolates in Brazil, the whole population of T. gondii isolates from chickens in Brazil and the population surveyed in this study were 0.97, 0.95 and 0.90, respectively. Seven of the 11 genotypes revealed from chicken isolates from MS are newly described genotypes and six of them each have a single isolate. In conclusion, the results obtained from isolates in MS corroborate previous studies on T. gondii isolates in Brazil, thus confirming their diversity and atypicality. Nonetheless, the applicability of PCR-RFLP markers for epidemiological inferences remains controversial.
Subject(s)
Chickens , Poultry Diseases/parasitology , Toxoplasma/genetics , Toxoplasmosis, Animal/parasitology , Animals , Genotype , Mice , Phylogeny , Poultry Diseases/epidemiology , Toxoplasma/pathogenicity , Toxoplasmosis, Animal/epidemiologyABSTRACT
Toxoplasma gondii is an aetiological agent of toxoplasmosis, which commonly causes diarrhoea in a number of species. This observation and the parasite's affinity for the nervous tissue support the theory that T. gondii infection may affect the myenteric neurons. The aim of this study was to evaluate the changes caused by T. gondii (genotype III) in the myenteric neurons of the jejunum in rats. Fifteen rats were distributed into three groups: control (CG), inoculated for 30 days (G30) and inoculated for 90 days (G90). Rats from the G30 and G90 groups received an oral inoculum with 500 oocysts from a genotype III (M7741) T. gondii strain. At 180 days of age, all animals were anaesthetised and euthanised. Whole mounts were stained by using Giemsa (total population) and NADPH-diaphorase (nitrergic subpopulation) histochemistry. Maintenance of the width, length, area and neuronal density was observed; there was neuronal atrophy in the G30 group and a tendency to hypertrophy in the G90 group. Rats inoculated orally with sporulated oocysts did not show clinical illness or macroscopic or microscopic lesions, as do the majority of animal species. Therefore, infection was confirmed by a serum agglutination test; 30 days of infection caused increased weight gain and atrophy of myenteric neurons. At 90 days post-infection, weight gain became normal, and myenteric neurons hypertrophied.
Subject(s)
Jejunum/pathology , Myenteric Plexus/pathology , Neurons/pathology , Toxoplasma , Toxoplasmosis, Animal/pathology , Animals , Chronic Disease , Jejunum/parasitology , Male , Myenteric Plexus/parasitology , Neurons/parasitology , Rats , Rats, WistarABSTRACT
Mycobacterium spp. and other pathogens were investigated in 258 swine lymph nodes (129 with and 129 without apparent lesions), and 120 lymph nodes (60 with and 60 without lesions) from wild boars (Sus scrofa). A total of lymph nodes from swine and wild boars were collected of different animals. Submaxillar and mesenteric lymph nodes were submitted to microbiological examination and colonies suggestive of Mycobacterium spp. (alcohol-acid bacilli) were submitted to PCR Restriction Assay (PRA). In swine with lymphadenitis, Mycobacterium spp. (24.1%) and Rhodococcus equi (13.2%) were the most prevalent microorganisms, while in lymph nodes without lesions were identified a complex of microorganisms, including of environmental mycobacteria. In wild boars with lymphadenitis, ß-haemolytic Streptococcus (10.0%), Mycobacterium spp (8.4%) and R. equi (6.6%) were the most frequent. Among mycobacterias were identified predominantly Mycobacterium avium subspecies type 1 (48.3%) and M. avium subspecies type 2 (16.1%), followed by Mycobacterium intracellulare, Mycobacterium szulgai,Mycobacterium fortuitum, Mycobacterium gordonae, Mycobacterium simiae, Mycobacterium nonchromogenicum and Mycobacterium intracellulare type 2.
Subject(s)
Gram-Positive Bacterial Infections/veterinary , Lymph Nodes/microbiology , Mycobacterium/isolation & purification , Rhodococcus equi/isolation & purification , Streptococcus/isolation & purification , Swine Diseases/microbiology , Animals , Gram-Positive Bacterial Infections/microbiology , SwineABSTRACT
The virulence genes and plasmid profiles of 23 Rhodococcus equi isolates from 258 lymph nodes from domestic pigs (129 nodes with lesions and 129 without lesions) and 120 lymph nodes from slaughtered wild boars (60 nodes with lesions and 60 without) were characterized. R. equi was obtained from 19 lymph nodes of domestic pigs, 17 with, and two without lesions, and from four lymph nodes with lesions, from wild boars. The 23 isolates were tested for the presence of vapA and vapB genes, responsible for the 15-17 and 20 kDa virulence-associated proteins, respectively, by PCR in order to characterize as virulent (VapA), intermediately virulent (VapB) and avirulent. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. Of the 19 domestic pigs strains, seven (36.8%) were avirulent and 12 (63.2%) were intermediately virulent, with the intermediately virulent isolates being plasmid types 8 (8 isolates), 10 (2 isolates), 1 (1 isolate) and 29 (1 isolate). The plasmid type of four strains isolated from wild boars was also intermediately virulent type 8. None of the domestic pigs and wild boar isolates showed the vapA gene. These findings demonstrate a high occurrence of plasmid type 8 in isolates from pigs and wild boars, and the similarity of plasmid types in the domestic pigs, wild boars and human isolates in Brazil.
Subject(s)
Actinomycetales Infections/veterinary , Plasmids/genetics , Rhodococcus equi/genetics , Rhodococcus equi/pathogenicity , Sus scrofa , Swine Diseases/microbiology , Actinomycetales Infections/epidemiology , Actinomycetales Infections/microbiology , Animals , Brazil/epidemiology , Swine , Swine Diseases/epidemiology , VirulenceABSTRACT
Toxoplasma gondii strains are genetically diverse in South America. To date, hundreds of T. gondii isolates from different animal hosts were genotyped in Brazil, most of them are different from those identified around the world. This study aimed to determine T. gondii infection rate in sheep from Brazilian slaughterhouses, as well as the genotype of these isolates. T. gondii antibodies were detected in 66/602 (10.96%) serum samples through modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT). MAT-HS and IFAT-IgG presented high concordance (0.95) and strong correlation (r=0.79). T. gondii DNA was detected in tissue samples of 33% (22/66) serum positive sheep by PCR of the 529 bp repetitive element. In the bioassay in mice, T. gondii were detected in mice brain or muscle tissues in 30% (20/66) of serum positive sheep. Positive samples were typed through Restriction Fragment Length Polymorphism (RFLP-PCR) using 11 markers: SAG1, SAG2 (5'-3'SAG2 and alt.SAG2), SAG3, BTUB, GRA6, L358, c22-8, c29-6, PK1, Apico and CS3. Of 22 samples, 13 were positive and 9 genotypes were identified. Four of these 9 genotypes are unique. Nine samples had negative results in RFLP-PCR typing, which may be due to low DNA concentration. Six isolates were virulent killing mice between 12 and 25 days postinfection. Two non-virulent isolates belonged to clonal type II genotype, which were not observed in Brazil previously. These findings confirm the high diversity and high frequency of virulent genotypes among Brazilian animals. This study also proved the presence of type II T. gondii in Brazil.
Subject(s)
Genotype , Sheep Diseases/parasitology , Toxoplasma/genetics , Toxoplasmosis, Animal/parasitology , Abattoirs , Animals , Antibodies, Protozoan/blood , Brazil/epidemiology , Gene Expression Profiling , Mice , Sheep , Sheep Diseases/blood , Sheep Diseases/epidemiology , Toxoplasmosis, Animal/blood , Toxoplasmosis, Animal/epidemiologyABSTRACT
Pigeons (Columba livia) cohabit with humans in urban and rural areas, representing a public health problem since microorganisms are transmitted through the inhalation of dust from their dry feces (chlamydiosis) and through ingestion of their undercooked or poorly refrigerated meat (toxoplasmosis). This study aimed to evaluate the presence of Chlamydophila psittaci and Toxoplasma gondii in pigeons from four cities in São Paulo State, Brazil. C. psittaci was evaluated through hemi-nested polymerase chain reaction (hnPCR) using cloacal and tracheal swabs, whereas T. gondii specific antibodies were assessed by means of modified agglutination test (MAT), mouse brain and muscle bioassay, and polymerase chain reaction (PCR). To confirm the infection in mice, T. gondii antibodies were assayed by using indirect fluorescent antibody test (IFAT). Considering C. psittaci, 40/238 (16.8%; 95%CI 12.6-22.1%) samples were positive according to hnPCR, especially for the cities of São Paulo (42.5%) and Bauru (35%). As regards T. gondii, 12/238 (5%; 95%CI 2.9-8.6%) serum samples were positive according to MAT. Of these, five samples had titer equal to 1:8; six samples, 1:16; and one sample, 1:32. Bioassay, IFAT and PCR were negative for mouse toxoplasmosis. The absence of T. gondii antibodies suggests that pigeons may be infected with a low concentration of the agent, not detected by the antigen test. Thus, C. psittaci represents an actual problem concerning bird health.
Subject(s)
Bird Diseases/parasitology , Columbidae , Psittacosis/epidemiology , Toxoplasma , Toxoplasmosis, Animal/epidemiology , Animals , Antibodies, Protozoan/blood , Bird Diseases/blood , Bird Diseases/epidemiology , Brazil/epidemiology , Chlamydophila psittaci , Mice , Psittacosis/blood , Toxoplasmosis, Animal/bloodABSTRACT
Toxoplasma gondii infection has been diagnosed in pigs all over the world. Economical losses are generally related to reproductive disorders. Toxoplasma infection is also a matter of public health because tissue cysts of the parasite may remain in pork and pork products, and become sources of human infection. The objective of this study was to evaluate the frequency and risk factors associated with Toxoplasma infection in certified and non-certified pig breeding farms in the Toledo microregion, in the State of Paraná, Brazil which includes the cities of Toledo, Nova Santa Rosa, Sao José das Palmeiras and Sao Pedro do Iguaçu. Relative frequency of infection was 13.4%, independently of the type of farm. Logistic regression analysis showed that the following factors were associated with infection: absence of workers exclusive for each area of the farm, access of other animals to feeders and drinkers, lack of lids in drinkers, lack of rodent control measures, mean piglet number and weight at weaning per female.