Main Causes of Death in Piglets from Different Brazilian Nursery Farms Based on Clinical, Microbiological, and Pathological Aspects.

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.

Evaluation of buffered Trypanosoma evansi antigen and rapid serum agglutination test (BA/Te) for the detection of anti-T. evansi antibodies in horses in Brazil.

Surra is an infectious disease caused by Trypanosoma evansi, which affects a large number of domestic and wild animal species. Infection control is based on rapid diagnosis followed by treatment of sick animals. This study aimed to evaluate a buffered T. evansi antigen and rapid serum agglutination test (BA/Te) for the detection of anti-T. evansi antibodies in serum samples of horses. For this purpose, 445 serum samples from horses were evaluated and the results compared with the diagnosis by CATT/T. evansi. Our data show a sensitivity of 92%, specificity of 91% and a degree of agreement kappa (κ) of 0.82 (95% CI: 0.771-0.877, P < 0.01) between BA/Te and CATT/T. evansi. Antigen specificity was also evaluated against reactive serum for other infectious agents circulating in equine herds. In conclusion, our findings show that BA/Te has the potential to be a practical and quick screening method for the detection of anti-T. evansi antibodies in horses.

First outbreak of autochthonous "surra" in horses in Santa Catarina State, Brazil: Parasitological, hematological and biochemical characteristics.

This study reports the first autochthonous "surra" outbreak in horses in the State of Santa Catarina, southern Brazil. Six horses with clinical suspicion of trypanosomosis had the natural infection by T. evansi confirmed by PCR and rapid serum agglutination test. Clinical, parasitological, and hematology evaluations were performed at initial observation (T0) and 90 days after (T1). At T0, all animals that tested positive for T. evansi in PCR presented with severe clinical signs and out of normal range hematological hematological (hematocrit, leukocytes, platelets, hemoglobin) and serum biochemical parameters (alanine aminotransferase (ALT), creatinine phosphokinase (CPK), lactate dehydrogenase (LDH), creatinine, bilirubin, and glucose). At T1, the progressive clinical recovery of animals, normalization of hemato-biochemical parameters, and negative PCR results for T. evansi were observed. It was not possible to identify the vector/mechanism of transmission through which animals were infected; therefore, the implementation of surveillance and control measures is essential to prevent the spread of this disease in horse herds, as well as to other animal species.

Risk factors related to seropositivity for Leptospira spp. in horses in the Serra Catarinense region / Fatores de risco relacionados a soropositividade para Leptospira spp. em equinos na região da Serra Catarinense

ABSTRACT: Leptospirosis is a zoonotic infectious disease caused by the bacterium Leptospira spp. The consequences of infection in horses are unclear, and the serogroups involved vary depending on the region. This study aimed to evaluate the seropositivity of unvaccinated horses in the Serra Catarinense region and the possible risk factors associated with the infection. We used 207 horses from 26 properties, with no clinical history of infection and unvaccinated for leptospirosis. We submitted serum samples to the microscopic agglutination serum (SAM) to obtain the main reference serogroups. We considered animals with titre ≥100 as infected. The results of the serology revealed seropositivity in 45.4% (94/207) of the animals studied. The most frequent antibodies reportedwere those against the serogroups Australis 16.9% (35/207), Ichterhaemorrhagiae 14.4% (30/207), and Grippotyphosa 5.31% (11/207). Among the primary risk factors associated with the infection we reported extensive rearing system (OR = 1.27; P<0.05) and the presence of other animal species such as cattle (OR = 3.85; P<0.01) and capybaras (OR = 2.07; P=0.06). The findings presented in this study showed that leptospirosis is endemic in horses in the Serra Catarinense region, and revealed the need for emergency surveillance and control measures specific to this important zoonosis.

RESUMO: A leptospirose é uma doença infecciosa zoonótica, causada pela bactéria Leptospira spp. Em equinos, as consequências da infecção não são claras e os sorogrupos envolvidos variam dependendo da região. Sendo assim, este trabalho objetivou avaliar a soropositividade de Leptospira spp. em equinos não vacinados na região serrana Catarinense e estimar os possíveis fatores de risco associados à infecção. Para tal, foram estudados 207 equinos, de 26 propriedades, sem histórico clínico da infecção e não vacinados para leptospirose. As amostras de soro foram submetidas ao ensaio de soro aglutinação microscópica (SAM) para os principais sorogrupos de referência. Animais com título ≥100 foram considerados reagentes. Os resultados da sorologia revelaram uma soropositividade de 45,4% (94/207) para os animais estudados. Com maior frequência foram observados anticorpos contra os sorogrupos Australis 16,9% (35/207), Icterohaemorrhagiae 14,4% (30/207), e Grippotyphosa 5,31% (11/207). Dentre os principais fatores de risco associados à infecção observados, foram o sistema de criação extensivo (OR=1,27; P<0,05) e a presença de outras espécies animais como bovinos (OR=3,85; P<0,01) e capivaras (OR=2,07; P=0,06). Os achados apresentados neste estudo mostraram que a leptospirose em equinos é endêmica na região da serra Catarinense, e revelam a necessidade de medidas emergenciais de vigilância e controle específicas para esta importante zoonose.

Molecular detection of Mycoplasma synoviae and avian reovirus infection in arthritis and tenosynovitis lesions of broiler and breeder chickens in Santa Catarina State, Brazil.

Infectious arthritis or tenosynovitis in broiler and breeder chickens results in major loss of productivity because of reduced growth and downgrading at processing plants. The most common causative agents of avian infectious arthritis are the bacterium Mycoplasma synoviae and avian reoviruses (ARVs) (family Reoviridae, genus Orthoreovirus). In this study, we evaluated the occurrence of these two pathogens in arthritis or tenosynovitis lesions of broilers and breeder flocks in southern Brazil using molecular detection. Tissue sections from tibiotarsal joints with visible lesions from 719 broilers and 505 breeders were analysed using pathogen-specific polymerase chain reaction (PCR) assays. In breeders, 41.2% (n = 296) of lesions were positive for M. synoviae, 26.4% (n = 190) were positive for ARV, while co-infection was present in 12.2% (n = 88) of the samples. In broilers, 20.8% (n = 105) of lesions were positive for M. synoviae, 11.9% (n = 60) for ARV and 7.7% (n = 39) of these cases were positive for both pathogens. Post-mortem examination revealed lesions with varying degrees of gross pathological severity. Histopathological examination showed intense, diffuse lymphohistiocytic inflammatory infiltrates with heterophil accumulation, primarily in the synovial capsule and digital flexor tendon, in all samples. Improved strategies for early detection and control of these major avian pathogens are highly desirable for preventing the spread of infection and reducing economic losses in the poultry industry.

Adipose-Derived Stem Cells Decrease Bone Morphogenetic Protein Type 2-Induced Inflammation In Vivo.

PURPOSE: Recombinant human bone morphogenetic protein type 2 (rhBMP-2) has been used to promote bone regeneration. In contrast, some reports have suggested rhBMP-2 does not provide advantages over autogenous bone grafting owing to the undesirable postoperative symptoms of this growth factor. Because the undesirable symptoms of rhBMP-2 are usually promoted by inflammation, this study evaluated the in vivo effect of human adipose-derived stem cells (ASCs) incorporated into polylactic co-glycolic acid (PLGA) scaffolds in decreasing the inflammatory response induced by a low dose of rhBMP-2. MATERIALS AND METHODS: PLGA scaffolds were characterized and loaded with rhBMP-2 1, 2.5, or 5 µg per scaffold (n = 6) and the in vitro released protein amounts were quantified at 7 hours and 1, 7, and 21 days after loading (n = 3). The muscle tissue of 6 beagles received the following treatments: PLGA, PLGA plus rhBMP-2 (2.5 µg), and PLGA plus rhBMP-2 plus ASCs (1 × 10(6) ASCs). The samples were evaluated 45 days after surgery. Statistical analyses were performed and the P value was set at .05. RESULTS: PLGA plus rhBMP-2 plus ASCs yielded the smallest number of inflammatory foci (P < .001) and giant cells (P < .001) and the largest number of angiogenesis sites (P < .001). CONCLUSIONS: Human ASCs administered in vivo into PLGA scaffolds with a low dose of rhBMP-2 decrease tissue inflammation and increase angiogenesis in muscular sites.

Adipose-derived stem cells incorporated into platelet-rich plasma improved bone regeneration and maturation in vivo.

BACKGROUND/AIM: Some cases of tooth loss related to dental trauma require bone-grafting procedures to improve the aesthetics before prosthetic rehabilitation or to enable the installation of dental implants. Bone regeneration is often a challenge and could be largely improved by mesenchymal stem cells therapy. However, the appropriate scaffold for these cells still a problem. This study evaluated the in vivo effect of human adipose-derived stem cells incorporated into autogenous platelet-rich plasma in bone regeneration and maturation. MATERIAL AND METHODS: Adipose-derived stem cells were isolated from lipoaspirate tissues and used at passage 4. Immunophenotyping and multilineage differentiation of cells were performed and mesenchymal stem cells characteristics confirmed. Bicortical bone defects (10 mm diameter) were created in the tibia of six beagle dogs to evaluate the effect of adipose-derived stem cells incorporated into platelet-rich plasma scaffolds, platelet-rich plasma alone, autogenous bone grafts, and clot. Samples were removed 6 weeks postsurgeries and analyzed by quantification of primary and secondary bone formation and granulation tissue. RESULTS: Adipose-derived stem cells incorporated into platelet-rich plasma scaffolds promoted the highest bone formation (primary + secondary bone) (P < 0.001), the highest bone maturation (secondary bone) (P < 0.001), and the lowest amount of granulation tissue (P < 0.001). CONCLUSIONS: Adipose-derived stem cells incorporated into platelet-rich plasma scaffolds promote more bone formation and maturation, and less granulation tissue in bone defects created in canine tibia. Therefore, platelet-rich plasma can be considered as a candidate scaffold for adipose-derived stem cells to promote bone regeneration.

Outbreak of autochthonous canine visceral leishmaniasis in Santa Catarina, Brazil / Surto autóctone de leishmaniose visceral canina no Estado de Santa Catarina

The present study reports the first outbreak of autochthonous canine visceral leishmaniasis in Florianópolis, Santa Catarina, southern Brazil. Following the report of two cases of CVL, the Control Center of Zoonotic Diseases conducted a serological survey by ELISA and IFAT assays in seven districts of the Santa Catarina Island. Eleven seropositive dogs of autochthonous transmission were used in the present study. Infection by Leishmania sp. was confirmed by parasitological examination of bone marrow, liver, spleen and lymph nodes, culture in Schneider's medium and PCR. Leishmania sp. isolates were characterized by PCR-RFLP and hybridization with specific probes, allowing for the identification of Leishmania infantum. Autochthonous transmission of this disease in an area with high tourist traffic presents a major public health concern and signifies the emergence of an important zoonosis in southern Brazil. Therefore, the implementation of surveillance and control measures is imperative to prevent the spread of the disease among the canine population as well as transmission to the human population.

O presente estudo relata o primeiro surto autóctone de leishmaniose visceral canina (LCV) em Florianópolis, Santa Catarina, Brasil. Durante levantamento soro-epidemiológico realizado pelo Centro de Controle de Doenças Zoonóticas (CCZ) envolvendo 2.124 cães, 29 (1,37%) foram soropositivos para VL (ELISA + RIFI). Onze cães positivos por transmissão autóctone foram utilizados no presente estudo. A confirmação da infecção por Leishmania sp. foi realizada pelo exame parasitológico da medula óssea, fígado, baço e linfonodos, cultura em meio Schneider e PCR. Os isolados de Leishmania sp. foram caracterizados por PCR-RFLP e hibridação com sondas específicas, permitindo a identificação de Leishmania infantum. A transmissão autóctone da LCV em uma área com grande fluxo turístico como Florianópolis representa um preocupante risco à saúde pública e o surgimento de uma importante zoonose no sul do Brasil. Neste contexto, a implementação de medidas de vigilância e controle da doença são fundamentais para evitar a propagação da doença entre a população canina, bem como a transmissão para a população humana.

Asymptomatic cattle naturally infected with Mycobacterium bovis present exacerbated tissue pathology and bacterial dissemination.

Rational discovery of novel immunodiagnostic and vaccine candidate antigens to control bovine tuberculosis (bTB) requires knowledge of disease immunopathogenesis. However, there remains a paucity of information on the Mycobacterium bovis-host immune interactions during the natural infection. Analysis of 247 naturally PPD+ M. bovis-infected cattle revealed that 92% (n = 228) of these animals were found to display no clinical signs, but presented severe as well as disseminated bTB-lesions at post-mortem examination. Moreover, dissemination of bTB-lesions positively correlated with both pathology severity score (Spearman r = 0.48; p<0.0001) and viable tissue bacterial loads (Spearman r = 0.58; p = 0.0001). Additionally, granuloma encapsulation negatively correlated with M. bovis growth as well as pathology severity, suggesting that encapsulation is an effective mechanism to control bacterial proliferation during natural infection. Moreover, multinucleated giant cell numbers were found to negatively correlate with bacterial counts (Spearman r = 0.25; p = 0.03) in lung granulomas. In contrast, neutrophil numbers in the granuloma were associated with increased M. bovis proliferation (Spearman r = 0.27; p = 0.021). Together, our findings suggest that encapsulation and multinucleated giant cells control M. bovis viability, whereas neutrophils may serve as a cellular biomarker of bacterial proliferation during natural infection. These data integrate host granuloma responses with mycobacterial dissemination and could provide useful immunopathological-based biomarkers of disease severity in natural infection with M. bovis, an important cattle pathogen.

Rapid detection of Mycoplasma synoviae and avian reovirus in clinical samples of poultry using multiplex PCR.

Mycoplasma synoviae and avian reovirus (ARV) are associated with several disease syndromes in poultry and cause notable global economic losses in the poultry industry. Rapid and efficient diagnostics for these avian pathogens are important not only for disease control but also for prevention of clinical disease progression. However, current diagnostic methods used for surveillance of these diseases in poultry flocks are laborious and time-consuming, and they have low sensitivity. The multiplex PCR (mPCR) developed in this study has been proven to be both sensitive and specific for simultaneous M. synoviae and ARV detection and identification in clinical samples. To evaluate the mPCR assay, the diagnostic test was applied to different clinical samples from natural and experimental M. synoviae and ARV-infected poultry. Results were compared with serologic, single PCR, and immunofluorescence analyses. Tibiotarsal articulation could be the best target for simultaneous detection of M. synoviae and ARV infection. The detection limit by visualization of mPCR-amplified products was 100 pg for both pathogens. Overall, the mPCR developed and standardized in this research is a useful tool for diagnosis and screening and for surveillance and control of M. synoviae and ARV infection in poultry flocks.

Características clínicas e anatomo-histopatologicas da infecção experimental mista por Orthoreovirus aviario e Mycoplasma synoviae em frangos de corte / Clinical and histologic lesions of mixed infection with Avian orthoreovirus and Mycoplasma synoviae in broilers

A artrite infecciosa em frangos de corte representa um problema sanitário e econômico de grande impacto, provocando perdas de produtividade e nos processos de produção e industrialização. Os principais agentes etiológicos associados aos casos de artrites e tenossinovites infecciosas em aves são Mycoplasma synoviae (MS) e Orthoreovirus aviario (ARV). Esse trabalho propôs investigar as alterações anatomohistopatológicas causadas pela infecção experimental concomitante por Mycoplasma synoviae e Orthoreovirus aviario em frangos de corte e confirmar a presença dos agentes através das técnicas de PCR e imuno-luorescência indireta (RIFI). Para tal foram utilizados 16 frangos de corte, alojados em cama, com fornecimento de ração e água ad libitum. A infecção experimental foi realizada utilizando amostras atenuadas de MS e de ARV. Clinicamente as aves inoculadas apresentaram apatia e edemaciação da região da articulação tíbiotársica. Após 30 dias procedeu-se a eutanásia e a necropsia das aves. Na análise histopatológica constatou-se o efeito da infecção mista com MS e ARV sobre os diferentes órgãos/tecidos. Todos os animais apresentaram quadro de artrite e tenossinovite caracterizado pela presença de infiltrado inflamatório linfohistiocitário difuso, com acúmulo de heterófilos na cápsula articular/membrana sinovial e tendão flexor digital. Além disso, foi possível observar infiltrado inflamatório na traquéia, nos pulmões e sacos aéreos, no fígado, baço, pericárdio e proventrículo. A utilização da RIFI foi necessária para visualizar a presença de ambos os agentes nas articulações, identificando a presença de antígenos do ARV e do MS. A técnica de PCR constatou positividade do MS na traquéia, pulmões/sacos aéreos, cápsula articular/membrana sinovial e liquido sinovial. Já para o ARV a PCR foi positiva em amostras de fígado, baço, cápsula articular/membrana sinovial e tendão flexor digital. Com base nas lesões observadas e nos dados da literatura, sugere-se a ação concomitante por MS e ARV nos diferentes tecidos.

Infectious arthritis in broiler represents an economic and health problem resulting in severe losses due to retarded growth and down grading at slaughterhouse. The most common agents associated with cases of infectious arthritis in poultry are Mycoplasma synoviae and Avian orthoreovirus. This study proposed to investigate the histopathological changes caused by mixed infection with Mycoplasma synoviae and Avian orthoreovirus in broilers and confirm the presence of the agents through PCR and immunofluorescence assay (IFA). We used 16 broiler chickens, housed in bed, with supply of food and water ad libitum. Ten-day-old broilers were infected with Mycoplasma synoviae and Avian orthoreovirus. Clinically, they showed lethargy and swelling of the hock joint. After 30 days, we proceeded to their euthanasia and necropsy. Histological lesions were observed due to the mixed infection with Mycoplasma synoviae and Avian orthoreovirus in different tissues. The histopathology of the joints was characterized by infiltration of heterophil leucocytes in the synovial membrane and the digital flexor tendon. The inflammatory process was also found in trachea, lungs, air sac, liver, spleen, pericardium and proventriculus. The use of IFA was necessary to verify the presence of both agents in the hock joints, identifying the antigens of Mycoplasma synoviae and Avian orthoreovirus. The presence of M. synoviae was detected by PCR in trachea, lung, air sacs, synovial membrane and synovial fluid. Avian orthoreovirus was detected with PCR in liver, spleen, synovial membrane and digital flexor tendon. In conclusion, this investigation suggests that a synergistic relationship exists between Mycoplasma synoviae and Avian orthoreovirus.

Selection of TcII Trypanosoma cruzi population following macrophage infection.

BACKGROUND: Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, which exhibits a high genetic variability. TcI, TcII, or mixed TcI/TcII strains may be found during acute human infection while mainly TcII parasites are present at the chronic stage of disease. In a previously studied Chagas disease outbreak, we identified mixed TcI/TcII strains in the vector Triatoma tibiamaculata and only TcII strains in infected humans, indicating that T. cruzi populations may be selected within the human host. METHODS: Utilizing molecular typing and cell biology techniques, we investigated the interaction of TcI, TcII, and mixed TcI/TcII strains with macrophages, an important cell population implicated in controlling protozoan infection. RESULTS: TcII but not TcI strains were selected by both human and murine macrophages in vitro and by peritoneal cavity cells in vivo. Biological analysis revealed that, compared with TcI, TcII strains display higher infective and multiplicative ability as well as lower doubling time inside macrophages. However, TcI and TcII strains present similar susceptibility to interferon-γ-activated macrophages in vitro. CONCLUSIONS: Taken together, our results reveal the existence of an intracellular selection process in macrophages that favors TcII, but not TcI, when infection occurs with vector-derived mixed TcI/TcII strains.

Simian recombinant adenovirus delivered by the mucosal route modulates gammadelta T cells from murine genital tract.

Although it has been demonstrated that mucosal immunization using vectors such as simian adenovirus (AdC) stimulates robust adaptive immune responses, there remains a paucity of information on the modulation of innate immune responses by such vectors. Using an established murine model of intravaginal immunization (Ivag), we have investigated whether mucosal gammadelta T cells participate in immune responses induced by AdC vectors. gammadelta T cell numbers were found to be increased in the vaginal tract. Moreover, gammadelta T cells isolated from the genital tract showed an activated phenotype and enhanced expression of cytokine gene. Altogether, our results demonstrate that AdC modulates gammadelta T cell responses and suggest that this cell population may influence immune responses following vaginal immunization.

Agentes bacterianos enteropatogênicos em suínos de diferentes faixas etárias e perfil de resistência a antimicrobianos de cepas de Escherichia coli e Salmonella spp / Enteropathogenic bacterial agents in pigs of different age groups and profile of resistance in strains of Escherichia coli and Salmonella spp. to antimicrobial agents

As enterites infecciosas bacterianas provocam severas perdas para a indústria suína em todo o mundo. Os objetivos deste trabalho foram determinar os agentes bacterianos, associados com a ocorrência de diarréia em suínos, em diferentes faixas etárias, no Estado de Santa Catarina, Brasil, e verificar o perfil de resistência das cepas de Escherichia coli e Salmonella spp, frente aos principais antimicrobianos utilizados em granjas de suínos. Os principais gêneros/espécies bacterianos diagnosticados foram Escherichia coli, Clostridium spp, Salmonella spp Brachyspira hyodysenteriae, Brachyspira pilosicoli e Lawsonia intracellularis. Os fatores de virulência de E. coli mais prevalentes na fase de maternidade foram F5 / (K99) 20 por cento, F6 / (987P) 16,3 por cento, F42 6,8 por cento e F41 5,7 por cento, já nas fases de creche e terminação, predominaram cepas com fimbrias F4 (K88) 11,2 por cento e 5,4 por cento, respectivamente. Para E. coli os maiores índices de resistência foram encontrados para oxitetraciclina (94 por cento) e tetraciclina (89,5 por cento) e os menores índices de resistência para neomicina (55 por cento), ceftiofur (57,4 por cento). Quanto às amostras de Salmonella spp, estas apresentaram maior resistência à oxitetraciclina (77 por cento), e à tetraciclina (42,1 por cento) e menor à gentamicina (3,5 por cento) e amoxicilina (4,8 por cento).

Infectious bacterial enteritis causes severe losses to the swine industry worldwide. The objective of this study was to determine the epidemiology of bacterial agents that are associated with the occurrence of diarrhea in pigs at different age groups, and to verify the profile of resistance of strains of Escherichia coli and Salmonella spp to the main antimicrobial agents. The main bacterial species diagnosed were Escherichia coli, Clostridium spp, Salmonella spp, Brachyspira hyodysenteriae, Brachyspira pilosicoli and Lawsonia intracellularis. The E. coli virulence factors of higher prevalence in preweaning piglets were F5 / (K99) 20 percent, F6 / (987P) 16.3 percent, F42 6.8 percent and F41 5.7 percent, whereas at the nursery and with finishing pigs, the prevalent strain was the fimbria F4 (K88) 11.2 percent e 5.4 percent, respectively. E. coli and Salmonella spp were highly resistant to oxytetracycline (94 percent) and tetracycline (90 percent), with the former having a low resistance to neomycin (55 percent) and ceftiofur (57 percent), and the latter to gentamicin (3.5 percent) and amoxicillin (4.8 percent).