Energy and mineral metabolism of peripartum mares and foals of the Quarter Horse breed.

BACKGROUND: Several studies have demonstrated variations in peripartum blood biochemical analytes used to monitor the health status of mares and their foals of different breeds. OBJECTIVES: We aimed to characterize the physiologic changes of peripartum energy and mineral metabolism in healthy Quarter Horse mares and their neonatal foals. METHODS: Blood samples were initially collected from 17 mares on days 60 and 30 prepartum, and from mares and their foals on the day of parturition (after colostrum ingestion), and on days 15, 30, and 60 postpartum. The serum concentrations of calcium, phosphorus, magnesium, total cholesterol (T-Cho), triglyceride, beta-hydroxybutyrate (BHBA), and nonesterified fatty acids (NEFA) were measured using commercial kits. Statistical differences were analyzed using one-way ANOVA followed by Tukey's test (parametric variables) or the Kruskal-Wallis test followed by Dunn's multiple comparison test (nonparametric variables). RESULTS: Fourteen of 17 healthy Quarter Horse mares and their neonatal foals remained in the study until termination. Serum BHBA, T-Cho, and calcium concentrations in mares showed significant differences during the observation period. Serum BHBA concentrations increased from 60 days prepartum to 60 days postpartum. Postpartum serum T-Cho and calcium concentrations significantly decreased until day-30 postpartum. In the foals, NEFA, T-Cho, calcium, and magnesium concentrations significantly differed between sampling times. In the foals, serum NEFA concentrations decreased from parturition until day-60 postpartum, while serum T-Cho, calcium, and magnesium concentrations were lowest at day-30 postpartum. CONCLUSIONS: The temporal changes observed in the blood biochemical analytes of this study could be used to improve the clinical evaluation of periparturient mare and neonatal foal Quarter Horses.

Severe outbreak of bovine neonatal diarrhea in a dairy calf rearing unit with multifactorial etiology.

This study describes the etiological diversity observed in a severe neonatal diarrhea outbreak with morbidity and mortality rates of 80 and 20%, respectively, with detection of mixed infections with viral, bacterial, and protozoan disease agents in a dairy calf rearing unit. Diarrheic fecal samples were collected from eight 5 to 18 days of age calves and were submitted to the investigation of the presence of rotavirus A (RVA), bovine coronavirus (BCoV), bovine kobuvirus (BKV), bovine viral diarrhea virus 1 and 2 (BVDV-1 and BVDV-2), enteropathogenic Escherichia coli (ETEC), Salmonella sp., and Cryptosporidium spp. Fragments of the small intestine of one calf with diarrhea that spontaneously died were submitted for histopathological analyses. The most frequent infectious agent detected in diarrheic fecal samples was BKV (8/8-100%), followed by RVA (5/8-62.5%), BVDV (5/8-62.5%), Cryptosporidium parvum (5/8-62.5%), ETEC (4/8-50%), and Cryptosporidium ryanae (1/8-12.5%). These etiological agents were found in mixed infections with two or more pathogens per diarrheic fecal sample. The association of viral and protozoan pathogens was the most frequently identified (37.5%) in these samples, followed by viral and bacterial (25%); viral, bacterial, and protozoan (25%); and only viral agents (12.5%). BCoV and Salmonella sp. were not identified in the diarrheic fecal samples analyzed. Additionally, histopathology of the small intestine diagnosed chronic lymphocytic enteritis. In conclusion, in calf rearing units, the adoption and strict monitoring of health management practices are critical to the success of this calf creation system.

Porcine lymphotropic herpesvirus DNA detection in multiple organs of pigs in Brazil.

We investigated the porcine lymphotropic herpesvirus (PLHV) DNA presence in multiple organs of pigs. Biological samples (n = 136) included tissue fragments of the central nervous system, heart, kidney, liver, lungs, spleen, urinary bladder, and urine. Sixty-eight (50%) organs were PLHV DNA-positive. None of the urine samples were detected with the virus genome. Although the presence of the PLHV DNA in the urinary bladder and kidney has been detected, it was not possible to show whether urine can be considered an effective route of virus shedding. This study warns to the risk of PLHV zoonotic transmission by xenotransplantation of tissues of porcine origin.

Risk factors associated with mammary tumors in female dogs / Fatores de risco para tumores mamários em cadelas de Uberlândia e Patos de Minas, Minas Gerais

Mammary tumors in female dogs are the most frequent and corresponds to half of the canine tumors. The objectives of this study were to determine the risk factors associated with the occurrence of mammary tumors in female dogs and to evaluate the macroscopic characteristics of these neoformations, using 386 dogs from the "Outubro Rosa Pets" events done within the cities of Uberlândia and Patos de Minas, Minas Gerais State, Brazil, in 2015 (n=194), 2016 (n=105) and 2017 (n=87). For the determination of risk factors, the binary logistic regression test (P<0.05) was performed. The occurrence of mammary tumors was 23.6% (91/386). The significant risk factors identified were increased age (P<0.001), overweight (P=0.048) and non-castration (P<0.001) with a chance of, respectively, 1.6, 2.3 and 9.3 times for the development of mammary tumors. In dogs with mammary tumors (n=91), 153 lesions were present, of which 39 female dogs had two or more lesions (42.8%). Most of the lesions were at the caudal abdominal (M4) and inguinal (M5) mammary glands (60.13%, 92/153). Relative to the size of the lesions, it was observed that in 78% of the female dogs the lesions were determined asT1 (<3cm), 16.5% were T2 (3-5cm) and 5.5% T3 (>5cm). At least 15.4% (14/91) of the dogs had one of the regional lymph nodes increased. In conclusion, the occurrence of mammary tumors in the evaluated population was 23.6% and that age, overweight and non-realization of ovariohysterectomy are risk factors associated with the development of mammary tumors.(AU)

Em cadelas os tumores mamários são os mais frequentes e correspondem a aproximadamente metade dos tumores em cães. Este estudo teve os objetivos de determinar os fatores de risco envolvidos na ocorrência de tumores mamários em cadelas e avaliar as características macroscópicas destas neoformações, utilizando 386 cadelas do evento "Outubro Rosa Pets" nos municípios de Uberlândia e Patos de Minas, Minas Gerais, Brasil, em 2015 (n=194), 2016 (n=105) e 2017 (n=87). Para a determinação dos fatores de risco utilizou-se o teste de Regressão logística binária (P<0,05). A ocorrência de tumores mamários foi de 23,6% (91/386). Os fatores de risco significativos identificados foram aumento da idade (P<0,001), sobrepeso (P=0,048) e não-castração (P<0,001) com a chance de, respectivamente, 1,6, 2,3 e 9,3 vezes de desenvolvimento de tumores mamários. Nas cadelas com tumores mamários (n=91), constatou-se a presença de 153 lesões, sendo que 39 cadelas apresentaram duas ou mais lesões (42,8%). A maioria das lesões localizaram-se nas mamas abdominais caudais (M4) e inguinais (M5) (60,13%; 92/153). Em relação ao tamanho das lesões, observou-se que 78% das cadelas eram T1 (<3cm), 16,5% T2 (3-5cm) e 5,5% T3 (>5cm). Pelo menos 15,4% (14/91) das cadelas apresentaram um dos linfonodos regionais aumentados. Conclui-se que a ocorrência dos tumores mamários na população avaliada foi de 23,6% e que a idade, sobrepeso e não ovariohisterectomia são fatores de risco para o desenvolvimento de tumores mamários.(AU)

Short communication: Proinflammatory gene expression relative to the collection technique of endometrial samples from cows with and without subclinical endometritis.

Uterine inflammation negatively affects reproductive performance and is an important cause of infertility and subfertility in dairy cows. Several studies have investigated the use of gene expression in endometrial samples collected by biopsy or cytology to evaluate the inflammatory response of the cow uterus. This study aimed to compare the expression of the CCL5, CXCL8, IL6, and IL1B genes in the bovine endometrium according to the site of sample collection [caruncular (C) or intercaruncular (IC)], the collection method (biopsy or cytology), and the category of inflammation based on endometrial cytology (zero, medium, or high) in subclinical endometritis. The reproductive tracts of dairy cows were collected from a slaughterhouse, and punch-biopsy samples of endometrial tissues were obtained from both regions (C and IC). Endometrial cells from these regions were collected with the cytobrush technique and then used for the analysis of mRNA expression by quantitative PCR. After counting polymorphonuclear cells (PMN) by endometrial cytology, 20 uteri with an ovary at stage I (d 1-4 of estrous cycle) were categorized into 3 groups. Uteri with 0% PMN (n = 10) were assigned to group zero, uteri with 5 to 15% PMN (n = 5) to group medium (12.2 ± 1.6% PMN), and uteri with >15% PMN (n = 5) to group high (53.8 ± 32.9% PMN). All data were analyzed with 2-way ANOVA with Bonferroni multiple comparison post test. The results from gene transcripts demonstrated that the region (C or IC) of the endometrial biopsy had no influence on any of the degrees of inflammatory reaction observed. However, gene expression was more elevated in the endometrium of cows with greater inflammation compared with those without inflammation (CCL5, CXCL8, IL6, IL1B) and those with medium inflammation (CCL5, IL6). Expression of the genes evaluated did not differ between the endometrium without inflammation and with medium inflammation. However, in the high inflammation group, all genes were comparatively more expressed in samples collected by cytology relative to those derived from biopsies for both anatomical regions. In conclusion, gene expression did not differ between the C and IC tissue. Samples collected from animals with greater inflammation had greater gene expression than those with zero or medium inflammation. In addition, cytology samples had greater gene expression than biopsy samples in the high inflammation group.

Rhodococcus equi pVAPN type causing pneumonia in a dog coinfected with canine morbillivirus (distemper virus) and Toxoplasma gondii.

Canine morbillivirus (previously, canine distemper virus, CDV) is a highly contagious infectious disease-causing agent that produces immunosuppressive infections and multiple clinical signs. Canine toxoplasmosis is an opportunistic disease characterized by enteric, pulmonary, and neuromuscular signs that might be confused with CDV-induced infections. Rhodococcus equi is a Gram-positive intracellular facultative bacterium that is also opportunistic in nature, and causes pyogranulomatous infections in humans and multiple host animals, although canine rhodococcosis is rare or unrecognized. The pathogenicity of R. equi is intimately related to the presence of plasmid-encoded virulence-associated proteins (Vap). Three host-adapted virulence plasmid types of R. equi have been recognized: the circular pVAPA and pVAPB are associated with equine and porcine strains, respectively, and the recently detected linear pVAPN virulence plasmid is related to bovine isolates. Nevertheless, data regarding the detection of host-adapted virulence plasmid types of R. equi isolated from companion animals are scarce. This report describes a case of an uncommon coinfection due to R. equi, T. gondii and CDV, which was diagnosed in a pet dog with respiratory distress. In this case, CDV most likely induced immunosuppression, which facilitated opportunistic infections by R. equi and T. gondii. The analysis of the virulence profile of R. equi revealed the novel pVAPN plasmid type, initially related to bovine strains. This is the second report of the bovine-associated pVAPN type in a pet dog, with an unusual coinfection with T. gondii and CDV. These findings represent a public health concern due to the close contact between pet animals and their owners, particularly because the pVAPN plasmid type was recently detected in people with HIV/AIDS from the same geographical region.

Pathologic, histochemical, and immunohistochemical findings in pulmonary and encephalitic cryptococcosis in a goat.

We describe the pathologic, histochemical, and immunohistochemical findings associated with pulmonary and encephalitic cryptococcosis in a 3-y-old, mixed-breed, nanny goat from central-west Brazil. The goat had progressive neurologic signs over 30 d; cryptococcosis was diagnosed antemortem by cytologic evaluation of cerebrospinal fluid. Treatment was initiated, but the animal died spontaneously shortly thereafter. Grossly, there was a large space-occupying gelatinous mass (cryptococcoma) in the left lung and smaller masses in the cerebral temporal and frontal cortex, thalamus, hippocampus, basal nuclei, and mesencephalon with consequent internal hydrocephalus. Histologic evaluation revealed marked granulomatous cryptococcal pneumonia and meningoencephalitis. Intralesional narrow-necked budding cryptococcal yeasts were identified on special stains (Mayer mucicarmine and Grocott methenamine silver) in sections of lung and brain. Immunohistochemistry utilizing a panel of monoclonal antibodies that selectively label capsules of Cryptococcus spp. was consistent with C. neoformans var. grubii.

Canine morbillivirus (canine distemper virus) with concomitant canine adenovirus, canine parvovirus-2, and Neospora caninum in puppies: a retrospective immunohistochemical study.

A retrospective immunohistochemical study was designed to investigate the frequency of concomitant traditional infectious disease pathogens in puppies that died suddenly and review the aspects of associated pathogenesis. Fifteen puppies were evaluated; the pathology reports and histopathologic slides of these animals were reviewed to determine the pattern of histopathologic lesions. The intralesional identification of antigens of canine (distemper) morbillivirus (CDV), canine adenovirus-1 and -2 (CAdV-1 and -2), canine parvovirus-2 (CPV-2), Toxoplasma gondii, and Neospora caninum was evaluated by IHC within the histopathologic patterns observed. All puppies contained CDV nucleic acid by molecular testing. The most frequent histopathologic patterns were intestinal crypt necrosis (n = 8), white matter cerebellar demyelination (n = 7), necrohaemorrhagic hepatitis (n = 7), interstitial pneumonia (n = 7), and gallbladder oedema (n = 5). All puppies contained intralesional antigens of CDV in multiple tissues resulting in singular (n = 3), and concomitant dual (n = 3), triple (n = 5) and quadruple (n = 4) infections by CAdV-1, and -2, CPV-2, and N. caninum; T. gondii was not identified. Concomitant infections by CDV was observed with N. caninum (100%; 1/1), CPV-2 (100%; 8/8), CAdV-1 (100%; 8/8), and CAdV-2 (100%; 8/8). Intralesional antigens of CDV and not CAdV-1 were identified in cases of gallbladder oedema. The "blue eye" phenomenon was histologically characterized by corneal oedema and degenerative lesions to the corneal epithelium, without inflammatory reactions.

Histophilus somni-associated syndromes in sheep from Southern Brazil

Abstract Histophilus somni is a Gram-negative bacterium that is associated with a disease complex (termed histophilosis) that can produce several clinical syndromes predominantly in cattle, but also in sheep. Histophilosis is well described in North America, Canada, and in some European countries. In Brazil, histophilosis has been described in cattle with respiratory, reproductive, and systemic disease, with only one case described in sheep. This report describes the occurrence of Histophilus somni-associated disease in sheep from Southern Brazil. Eight sheep with different clinical manifestations from five farms were investigated by a combination of pathological and molecular diagnostic methods to identify additional cases of histophilosis in sheep from Brazil. The principal pathological lesions were thrombotic meningoencephalitis, fibrinous bronchopneumonia, pulmonary abscesses, and necrotizing myocarditis. The main clinical syndromes associated with H. somni were thrombotic meningoencephalitis (n = 4), septicemia (n = 4), bronchopneumonia (n = 4), and myocarditis (n = 3). H. somni DNA was amplified from multiple tissues of all sheep with clinical syndromes of histophilosis; sequencing confirmed the PCR results. Further, PCR assays to detect Pasteurella multocida and Mannheimia haemolytica were negative. These findings confirmed the participation of H. somni in the clinical syndromes investigated during this study, and adds to the previous report of histophilosis in sheep from Brazil.

Diagnóstico diferencial das doenças neurológicas dos bovinos no estado do Paraná / Differential diagnosis of neurologic diseases of cattle in the state of Paraná

O objetivo deste trabalho foi identificar as doenças neurológicas que acometeram bovinos no estado do Paraná entre os anos de 2009 e 2015. A investigação aconteceu, preferencialmente, nas propriedades rurais onde os casos ocorreram. Foram registradas as informações sobre a evolução das doenças nos bovinos afetados do rebanho, e os prováveis fatores de risco foram identificados. Todos os procedimentos de exame físico geral e neurológico foram realizados sistematicamente para a caracterização da síndrome neurológica presente. Amostras de sangue e de líquor foram colhidas para a realização de exames laboratoriais. De acordo com o tempo de evolução e com a gravidade dos sinais clínicos observados, os bovinos doentes eram mantidos vivos para acompanhamento da evolução ou da resposta ao tratamento, ou eram submetidos à eutanásia seguida de necropsia. Fragmentos do sistema nervoso e dos demais órgãos foram colhidos para exame histopatológico. O exame de imunofluorescência direta e a prova biológica em camundongos foram realizados em todos os bovinos que morreram, com a finalidade de confirmar ou descartar o diagnóstico de raiva. Métodos laboratoriais específicos das rotinas de virologia, bacteriologia e toxicologia foram empregados, como complementares, para o estabelecimento do diagnóstico diferencial. Foram investigados 236 bovinos com doença neurológica, sendo 85 casos de ocorrência individual e 151 casos distribuídos por surtos que ocorreram em 79 rebanhos. As encefalopatias (180/236; 76,2%) predominaram sobre as mielopatias (27/236; 11,4%). As doenças inflamatórias determinadas por infecções (98/236; 41,5%) e as doenças tóxicas (91/236; 38,6%) foram as principais, enquanto as causas degenerativas (10/236; 4,2%), metabólicas (9/236; 3,8%), físicas (9/236; 3,8%), neoplásicas (4/236; 1,7%), e os defeitos congênitos (1/236; 0,4%) ocorreram menos frequentemente. Os casos inconclusivos somaram 5,9% (14/236). A meningoencefalite por BoHV-5 e a raiva foram as doenças de frequência maior e podem ser consideradas as mais importantes. Dentre as causas tóxicas, as intoxicações por plantas se destacaram (63/91; 69,2%) e foram responsáveis por 26,6% de todos os casos. Destacaram-se ainda a polioencefalomalácia, a meningoencefalite trombótica por Histophilus somni e o botulismo. Essas informações contribuem para que os médicos veterinários adotem condutas mais efetivas de diagnóstico e de prevenção, e são valiosas para o sistema oficial de vigilância epidemiológica do estado.(AU)

The aim of this study was to identify the neurological diseases that affected cattle in Paraná state between the years 2009 and 2015. The investigation took place, preferably, in the farms where cases occurred. Information on the evolution of the diseases in the affected cattle of the herd was recorded, and the probable risk factors were identified. All general and neurological examination procedures were performed systematically for the characterization of the neurological syndrome in each case. Samples of blood and CSF for laboratory exams were also collected. According to the evolution features and the severity of the observed clinical signs, the diseased cattle were kept alive to follow the progress of the disease, or were submitted to euthanasia followed by necropsy. Fragments of tissues from nervous system and other organs were collected for histopathological examination. Direct immunofluorescence test and biological test were performed on all the cattle that died, in order to confirm or rule out the diagnosis of rabies. Specific virology, bacteriology and toxicology laboratory methods were used as complementary exams in order to establish differential diagnosis. A total of 236 cattle with neurological disease were investigated, 85 cases of individual occurrence and 151 cases distributed by outbreaks that occurred in 79 herds. Encephalopathies (180/236, 76.2%) predominated over mielopathies (27/236, 11.4%). Inflammatory diseases caused by infections (98/236, 41.5%) and the toxic diseases (91/236, 38.6%) were the main causes, while degenerative (10/236, 4.2%), metabolic (9/236; 3.8%), physical (9/236, 3.8%), neoplastic (4/236, 1.7%), and congenital defects (1/236, 0.4%) occurred less often. The inconclusive cases were 5.9% (14/236). BoHV-5 meningoencephalitis and rabies were diseases of higher frequency and may be considered the most important. Among the toxic causes, plant poisonings were highlighted (63/91, 69.2%) and were responsible for 26.6% of all cases. Polioencephalomalacia, thrombotic meningoencephalitis caused by Histophilus somni and botulism were also highlighted. This information helps veterinarians to adopt more effective diagnostic and preventive approaches and is valuable to the state's official epidemiological surveillance system.(AU)

Histophilus somni-associated syndromes in sheep from Southern Brazil.

Histophilus somni is a Gram-negative bacterium that is associated with a disease complex (termed histophilosis) that can produce several clinical syndromes predominantly in cattle, but also in sheep. Histophilosis is well described in North America, Canada, and in some European countries. In Brazil, histophilosis has been described in cattle with respiratory, reproductive, and systemic disease, with only one case described in sheep. This report describes the occurrence of Histophilus somni-associated disease in sheep from Southern Brazil. Eight sheep with different clinical manifestations from five farms were investigated by a combination of pathological and molecular diagnostic methods to identify additional cases of histophilosis in sheep from Brazil. The principal pathological lesions were thrombotic meningoencephalitis, fibrinous bronchopneumonia, pulmonary abscesses, and necrotizing myocarditis. The main clinical syndromes associated with H. somni were thrombotic meningoencephalitis (n=4), septicemia (n=4), bronchopneumonia (n=4), and myocarditis (n=3). H. somni DNA was amplified from multiple tissues of all sheep with clinical syndromes of histophilosis; sequencing confirmed the PCR results. Further, PCR assays to detect Pasteurella multocida and Mannheimia haemolytica were negative. These findings confirmed the participation of H. somni in the clinical syndromes investigated during this study, and adds to the previous report of histophilosis in sheep from Brazil.

Molecular survey of infectious agents associated with bovine respiratory disease in a beef cattle feedlot in southern Brazil.

We investigated the occurrence of infectious pathogens during an outbreak of bovine respiratory disease (BRD) in a beef cattle feedlot in southern Brazil that has a high risk of developing BRD. Nasopharyngeal swabs were randomly collected from steers ( n = 23) and assessed for the presence of infectious agents of BRD by PCR and/or RT-PCR assays. These included: Histophilus somni, Mannheimia haemolytica, Pasteurella multocida, Mycoplasma bovis, bovine respiratory syncytial virus (BRSV), bovine coronavirus (BCoV), bovine viral diarrhea virus (BVDV), bovine alphaherpesvirus 1 (BoHV-1), and bovine parainfluenza virus 3 (BPIV-3). Pulmonary sections of one steer that died with clinical BRD were submitted for pathology and molecular testing. The frequencies of the pathogens identified from the nasopharyngeal swabs were: H. somni 39% (9 of 23), BRSV 35% (8 of 23), BCoV 22% (5 of 23), and M. haemolytica 13% (3 of 23). PCR or RT-PCR assays did not identify P. multocida, M. bovis, BoHV-1, BVDV, or BPIV-3 from the nasopharyngeal swabs. Single and concomitant associations of infectious agents of BRD were identified. Fibrinous bronchopneumonia was diagnosed in one steer that died; samples were positive for H. somni and M. haemolytica by PCR. H. somni, BRSV, and BCoV are important disease pathogens of BRD in feedlot cattle in Brazil, but H. somni and BCoV are probably under-reported.

Histopathological, immunohistochemical, and ultrastructural evidence of spontaneous Senecavirus A-induced lesions at the choroid plexus of newborn piglets.

Epidemic Transient Neonatal Losses (ETNL) is a disease of piglets caused by Senecavirus A (SVA) in which the method of dissemination and associated lesions are not well-defined. This study investigated the possible SVA-induced lesions by examining spontaneous infections in newborn piglets. Histopathology revealed ballooning degeneration of transitional epithelium, nonsuppurative meningoencephalitis, plexus choroiditis, and atrophic enteritis. RT-PCR identified SVA in all tissues evaluated and sequencing confirmed these results. Positive immunoreactivity to SVA was observed in endothelial and epithelial tissues of all organs evaluated. Semithin analysis revealed vacuolization of apical enterocytes of the small intestine, balloon degeneration and necrosis of endothelial cells of the choroid plexus (CP) and nonsuppurative choroid plexitis. Ultrathin evaluation demonstrated hydropic degeneration of apical enterocytes, degeneration and necrosis of endothelium of CP fenestrated capillaries, degeneration of ependymocytes associated with intralesional viral particles. It is proposed that SVA initially infects apical enterocytes of newborn piglets and probably enters the circulatory system with entry to the brain via the CP, by first producing an initial inflammatory reaction, with subsequent encephalitic dissemination. Consequently, SVA probably uses an enteric-neurological method of dissemination.

Complete Genome Sequence of Streptococcus iniae UEL-Si1, Isolated in Diseased Nile Tilapia (Oreochromis niloticus) from Northern Paraná, Southern Brazil.

The Streptococcus iniae UEL-Si1 strain was isolated from diseased Nile tilapia within the Paranapanema River Basin, Northern Paraná, Brazil. This is an emerging infectious disease agent of fish from Brazil, and sequencing of the complete genome is fundamental to understanding aspects relative to pathogenesis, infection, epidemiology, and immunity.

Clinical Manifestations of Senecavirus A Infection in Neonatal Pigs, Brazil, 2015.

We identified new clinical manifestations associated with Senecavirus A infection in neonatal piglets in Brazil in 2015. Immunohistochemical and molecular findings confirmed the association of Senecavirus A with these unusual clinical signs and more deaths. Other possible disease agents investigated were not associated with these illnesses.

Caracterização molecular de DNA de Delta papillomavirus bovino (BPV1, 2 e 13) em sarcoides equinos / Molecular characterization of bovine Deltapapillomavirus (BPV1, 2, and 13) DNA in equine sarcoids

Sarcoides são tumores fibroblásticos, considerados os tumores de pele mais comuns em pele de equinos e que raramente apresentam regressão espontânea. Papilomavírus bovino (BPV) tipos 1 e 2 são relacionados com a patogenia do sarcoide e, provavelmente, o BPV tipo 13 (BPV13), recentemente descrito, também pode estar associado com a formação dessa lesão. Neste estudo, 20 amostras de lesões cutâneas, sendo 12 constituídas por tecidos frescos e 8 amostras de tecido fixado em formalina e embebido em parafina, provenientes de 15 cavalos foram utilizadas para a identificação do DNA de BPV. A análise histopatológica (HE) confirmou todas as lesões como sarcoide. Para a amplificação do DNA de papilomavírus (PV) foram realizadas três reações de PCR. Como triagem, os primers IFNR2/IDNT2 foram utilizados para amplificar um fragmento da ORF L1 do PV. O segundo par de primersutilizado é complementar a sequência dos genes E5 e L2 de BPVs 1, 2 e 13. O terceiro par de primers(FAP59/FAP64) utilizado tem o gene L1 como alvo. A primeira e a segunda PCRs permitiram amplificar produtos em todas as amostras avaliadas. Entretanto, na terceira reação, na qual foram utilizados os primers FAP, foi possível amplificar produtos com tamanho molecular esperado somente nas amostras constituídas por tecidos frescos. O sequenciamento de nucleotídeos e as análises filogenéticas realizadas nos fragmentos E5L2 resultaram na identificação de BPV1, 2 e 13 em 14 (70%), 2 (10%) e em 4 (20%) amostras de sarcoides, respectivamente. As amostras de sarcoides de um dos animais continha somente o DNA de BPV1. Entretanto, nas amostras provenientes do segundo cavalo foi possível identificar o DNA de três tipos de Deltapapillomavirus bovino (BPV1, 2 e 13) em lesões distintas. Este estudo ratifica a presença do DNA de BPV1, 2 e 13 em lesões de sarcoides em equinos, além de identificar três tipos de BPVs em um mesmo animal e descrever pela primeira vez no Brasil a presença de BPV1 e 2 nesse tipo de lesão.

Sarcoids are fibroblastic lesions, which are considered as the most common skin tumors of horses; spontaneous regression rarely occurs. The bovine papillomavirus (BPV) types 1 and 2 may be involved in the pathogenesis of sarcoids, and probably the recently described BPV type (BPV13) might be associated with the pathogenesis of this lesion. This study characterized the DNA of BPVs in sarcoids from 15 horses from Brazil by analyzing 20 cutaneous lesions (12 recently collected; 8 from formalin-fixed paraffin-embedded (FFPE) tissues). Histopathology confirmed the proliferative lesions as sarcoids. Three PCRs were performed to amplify papillomavirus (PV) DNA. For screening, the primers IFNR2/IDNT2 were used to amplify a fragment of the PV L1 ORF. The second primer set was complementary to a common sequence of the E5L2 genomic region of BPV1, 2, and 13. The third primer pair (FAP59/FAP64) targeted a fragment of the PVs L1 ORF. The screening and E5L2 PCRs yielded amplicons in all samples evaluated. The FAP amplicons identified BPV1, 2, and 13 only from fresh tissue samples. The phylogenetic analyses of E5L2 resulted in the identification of BPV1, 2, and 13 in 14 (70%), 2 (10%), and 4 (20%) sarcoids, respectively. Two horses demonstrated multiple lesions: the sarcoids of one of these contained only BPV1 DNA and those of the other contained three types of bovine Deltapapillomavirus (BPV1, 2, and 13). This study confirmed the presence of BPV1, 2, and 13 DNA in equine sarcoids. Moreover, these findings represent the first description of three types of BPV diagnosed in the same horse, as well as the first confirmation of BPV1 and 2 in horses from Brazil.

Histophilus somni-induced thrombotic meningoencephalitis in cattle from northern Paraná, Brazil / Meningoencefalite trombótica-induzida por Histophilus somni em bovinos da região norte do Paraná

Thrombotic meningoencephalitis (TME) is a fatal neurological disease of cattle, predominantly from North America, that is caused by Histophilus somni with sporadic descriptions from other countries. This manuscript describes the occurrence of spontaneous TME in cattle from northern Paraná, Brazil. Most cattle had acute neurological manifestations characteristic of brain dysfunction. Hematological and cerebrospinal fluid analyses were not suggestive of bacterial infections of the brain. Histopathology revealed meningoencephalitis with vasculitis and thrombosis of small vessels that contained discrete neutrophilic and/or lymphocytic infiltrates admixed with fibrin at the brainstem, cerebral cortex, and trigeminal nerve ganglion of all animals. All tissues from the central nervous system used during this study were previously characterized as negative for rabies virus by the direct immunofluorescence assay. PCR and RT-PCR assays investigated the participation of infectious agents associated with bovine neurological disease by targeting specific genes of H. somni, Listeria monocytogenes, bovine herpesvirus -1 and -5, bovine viral diarrhea virus, and ovine herpesvirus-2. PCR and subsequent sequencing resulted in partial fragments of the 16S rRNA gene of H. somni from brain sections of all animals with histopathological diagnosis of TME; all other PCR/RT-PCR assays were negative. These findings confirmed the participation of H. somni in the neuropathological disease observed in these animals, extend the geographical distribution of this disease, and support previous findings of H. somni from Brazil.(AU)

Meningoencefalite trombótica (Thrombotic meningoencephalitis- TME) é uma doença neurológica fatal de bovinos ocasionada por Histophilus somni. A infecção tem sido descrita predominantemente na América do Norte e de forma esporádica em outros países. O objetivo deste estudo é relatar a ocorrência de TME em bovinos da região norte do estado do Paraná, Brasil. A maioria dos animais apresentaram sinais clínicos neurológicos característicos de disfunção cerebral aguda. Análises hematológicas e do fluido cerebrospinal não foram sugestivas de infecção bacteriana do cérebro. A histopatologia revelou meningoencefalite com vasculite e trombose de pequenos vasos com discreto infiltrado neutrofílico e/ou linfocítico mesclada com fibrina no tronco e córtex cerebral e no gânglio do nervo trigêmio de todos os animais. As amostras de sistema nervoso central incluídas nesse estudo foram previamente caracterizadas como negativas para raiva por meio de técnica de imunofluorescência direta. A participação de agentes infecciosos associados à doença neurológica em bovinos foi avaliada por técnicas moleculares como PCR e RT-PCR para amplificação parcial de genes de H. somni, Listeria monocytogenes, herpesvírus bovino 1 e 5, vírus da diarreia viral bovina e herpesvírus ovino 2. As seções do cérebro de todos os animais com diagnóstico histopatológico de TME foram positivas em PCR para a detecção do gene 16S rRNA de H. somni. O sequenciamento dos produtos amplificados confirmou a presença de DNA de H. somni nos fragmentos de cérebro avaliados. As reações de PCR/RT-PCR para todos os outros micro-organismos avaliados resultaram negativas. Os resultados desse estudo confirmaram a participação do H. somni nos episódios de doença neurológica observada nos animais avaliados, amplia a distribuição geográfica da TME e ratifica estudos prévios realizados no Brasil que demonstraram a presença de H. somni em outras formas de manifestação clínica das infecções por essa bactéria.(AU)

Identification of canine papillomavirus type 1 (CPV1) DNA in dogs with cutaneous papillomatosis / Identificação do DNA de Papillomavirus canino tipo 1 em cães com papilomas cutâneos no Brasil

Canine oral papillomavirus (COPV), also known as Canine Papillomavirus type 1 (CPV1), induces papillomas at the mucous membranes of the oral cavity and at the haired skin of dogs. The classification of Papillomavirus (PV) types is based on the L1 capsid protein and nucleotide sequence; so far, 14 CPV types have been described in several countries, but the molecular characterization of CPV in Brazil is lacking. This study investigated the presence of the PV in seven papillomas from four mixed breed dogs from Londrina/PR, Southern Brazil, by partial sequencing of the L1 gene. Seven exophytic cutaneous lesions were surgically removed and processed for histopathological and molecular characterization. Histopathology confirmed the lesions as viral papillomas due to typical histological features. Polymerase Chain Reaction (PCR) assay using the FAP59 and FAP64 primers targeted the L1 gene followed by sequence analysis of the amplicons identified CPV1 in all evaluated papilloma samples. This study represents the first description of CPV1 DNA associated with canine papillomatosis in Brazil.

O papilomavírus oral canino (COPV), também denominado Papillomavirus canino tipo 1 (CPV1), tem a capacidade de induzir papilomas na mucosa da cavidade oral e também em pele de cães. A classificação dos tipos de papilomavírus (PV) é baseada na proteína L1 do capsídeo e na sequência de nucleotídeos que a codifica. Atualmente são descritos 14 tipos de CPV, no entanto, ainda faltam estudos moleculares relacionados à identificação dos tipos de CPV no Brasil. O objetivo deste estudo foi investigar a presença de PV em fragmentos de papilomas obtidos de quatro cães sem raça definida, provenientes de Londrina/PR, região sul do Brasil, e definir o tipo viral por meio da análise da sequência parcial de nucleotídeos do gene L1. Sete lesões cutâneas foram cirurgicamente removidas e processadas ​​para a caracterização histopatológica e molecular. O exame histopatológico confirmou as lesões como papilomas. Foi realizada reação em cadeia de polimerase (PCR), utilizando os primers FAP59 FAP64 para a amplificação parcial do gene L1, seguida por análise das sequências dos produtos amplificados, que confirmou a presença do CPV1 em todas as amostras avaliadas. Este estudo representa a primeira identificação do DNA de CPV1 associado com papilomatose canina no Brasil.

Phaeohyphomycoses in a free-ranging loggerhead turtle (Caretta caretta) from Southern Brazil.

This report describes the occurrence of mycotic infection in a loggerhead turtle, Caretta caretta, found on Mostardas beach in the state of Rio Grande do Sul, Southern Brazil. The specimen was observed alive, emaciated, and died the following day. A necropsy was performed soon after death and tissue samples routinely processed for histopathological and molecular evaluation. Significant pathological alterations included multifocal to coalescing, 0.5-4 cm in diameter nodules were observed throughout the peritoneum and kidneys that revealed caseous, grayish content when sectioned; histopathological evaluation revealed severe peritonitis and nephritis associated with intralesional fungi. Fungal PCR that targeted the internal transcribed spacer region of fungi revealed three different species of fungi: Cladosporium cladosporioides and Alternata arborescens within the kidneys while Ampelomyces sp. was identified within peritoneal granulomas. C. cladosporioides and A. arborescens are melanized fungi that produce phaeohyphomycosis in a wide range of species. However, the importance of the identification of the mycoparasite Ampelomyces sp. DNA within the peritoneal granulomas remains unclear.