Proteinograma e teores de cobre, ferro e zinco no soro sanguíneo de ovelhas da raça Santa Inês com mastite experimental por Staphylococcus aureus

Costa N.A., Simao L.C.V., Santos R.A., Afonso J.A.B., Fagliari J.J., Cardoso E.C., Soares P.C. & Mendonca C.L. 2010. [Proteinogram and serum concentrations of copper, iron and zinc in Santa Ines ewes with Staphylococcus aureus experimentally induced mastitis.] Proteinograma e teores de cobre, ferro e zinco no soro sanguineo de ovelhas da raca Santa Ines com mastite experimental por Staphylococcus aureus. Pesquisa Veterinaria Brasileira 30(5):435-442. Clinica de Bovinos, Campus Garanhuns, Universidade Federal Rural de Pernambuco (UFRPE), Avenida Bom Pastor s/n, Boa Vista, Caixa Postal 152, Garanhuns, PE 55292-901, Brazil. E-mail: carlalopes.mendonca@gmail.comThe aim of the present study was to evaluate the effect of Staphylococcus aureus experimentally induced mastitis on proteinogram and serum concentrations of cupper, iron and zinc levels of Santa Ines primiparous ewes. The right mammary gland of ten healthy ewes was inoculated with 1,0x10(4) UFC/mL of S. aureus. Clinical examination and determination of serum concentrations of proteins by electrophoresis in polyacrylamide gel (SDS-PAGE), cupper, iron and zinc, as well plasma level of fibrinogen were measured before the inoculation (control) and 12h, 24h, 36h, 48h, 60h, 72h, 84h, 96h, 108h, 120h, 132h, 168h, 180h, 288h and 336h after bacteria inoculation. All animals experimentally infected presented clinical mastitis and subsequent loss of mammary gland function. The electrophoretogram allowed the identification of 23 proteins with molecular weights (MW) ranged from 26.000 to 185.000 daltons (Da) including acute-phase proteins, IgG and IgA. A significant increase (P<0,05) in haptoglobin, ceruloplasmin, IgG and IgA concentrations was observed. Antitrypsin and acid glicoprotein concentrations did not alter. The levels of iron and zinc decreased and the cupper concentration increased. A positive correlation between plasma fibrinogen and serum ceruloplasmin (r=0.74), haptoglobin (r=0.62) and IgA (r=0.62) was also identified. Results showed the importance of ceruloplasmin and haptoglobin as acute-phase proteins in ewes with intramammary infections and confirms fibrinogen as an inflammatory marker because its high correlation with specific proteins. The alterations in the serum levels of Cu, Fe and Zn suggest the action of inflammatory mediators triggered by S. aureus.

Breed and season influence on milk quality parameters and in mastitis occurrence

Os objetivos do presente estudo foram avaliar a performance das raças Jersey e Holandesa em diferentes condições de pluviosidade (estações seca e chuvosa) através do monitoramento de aspectos relacionados com a mastite subclínica (contagem de células somáticas, isolamento microbiológico, tipo de patógeno isolado), qualidade do leite (lactose, proteína, gordura e sólidos totais) e produção (media da produção de leite) de ambas as raças. O estudo foi conduzido em propriedade leiteira localizada no estado de São Paulo, Brasil. Oito visitas foram realizadas a propriedade, quatro em período de alta pluviosidade e outro em período de baixa pluviosidade. Amostras de leite foram colhidas de todos os animais para contagem eletrônica de células somáticas e determinação dos principais componentes do leite (proteína, gordura, sólidos totais, lactose). Gordura, proteína, sólidos totais e produção leiteira foram influenciadas tanto pela raça como pela estação, apresentando tendências similares para ambas raças em ambas estações. A contagem de células somáticas (CCS) apresentou resultados similares para ambas as raças. Vacas holandesas com infecção intramamária (IIM) apresentaram um maior aumento na CCS quando comparadas as vacas Jersey (P<0.001). Na estação seca, 53 animais tiveram IIM em pelo menos um mês do estudo. Destes, 32 eram da raça Holandesa e 21 da raça Jersey. Na estação chuvosa, 65 animais possuíram infecção intramamária, dos quais 43 eram da raça Holandesa e 22 da raça Jersey. A frequência de casos de IIM foi maior na estação chuvosa em comparação à estação seca. Vacas Jersey apresentaram uma menor chance de desenvolver sinais e sintomas de IIM em relação às vacas holandesas na estação chuvosa (razão de chances=0.52). O maior numero de casos de IIM na estação chuvosa pode ter contribuído para uma menor taxa de lactose no leite para ambas as raças, sendo que esta pode ser considerada como um indicador do status de IIM. Existiu uma prevalência de patógenos contagiosos durante todo o experimento. O modelo estatístico aplicado mostrou que patógenos ambientais foram mais frequentemente isolados da raça Jersey, independentemente do período do estudo. Aparentemente existem diferenças na resposta imune das raças Jersey e Holandesa.

Relationship between teat-end condition, udder cleanliness and bovine subclinical mastitis

The aims of the present study were to relate intramammary infection (IMI) occurrence and somatic cell count (SCC) with teat-end condition (TEC) and udder cleanliness (UC). Milk samples from 1931 teats were evaluated according to the presence of IMI and SCC. Scores were applied to teats according to the TEC and to UC. Teats ends with a very rough ring had the largest number of IMI when compared to the other three categories, as well as animals with dirtier udders. The change in a TEC score increased by around 30% the chance of IMI. Also, the chance of the animal developing IMI increased by approximately 47% when the UC score increased. No significant association between both scores and quarter SCC was found. It can be concluded that animals with very rough teat end rings and very dirty udders have a greater predisposition to IMI. (C) 2011 Elsevier Ltd. All rights reserved.

Molecular characterization and clonal diversity of methicillin-susceptible Staphylococcus aureus in milk of cows with mastitis in Brazil

Mastitis is an important disease for the dairy industry worldwide, causing economic losses and reducing milk quality and production. Staphylococcus aureus is a worldwide agent of this intramammary infection, which also causes foodborne diseases. The objective of this study was to determine the frequency of methicillin-susceptible Staphylococcus aureus (MSSA) isolates in milk of mastitis cows in Brazil and to analyze the genetic lineages and the content of antimicrobial resistance genes and virulence factors among these isolates. Fifty-six MSSA isolates were recovered from 1,484 milk samples (positive for the California mastitis test) of 518 cows from 11 different farms in Brazil (representing 51% of total Staph. aureus obtained), and they were further characterized. Methicillin-susceptible Staphylococcus aureus were isolated from 3.7% of California mastitis test-positive tested milk samples and from 6.2% of tested mastitic cows. Methicillin-susceptible Staphylococcus aureus isolates were characterized by spa typing, agr typing, and multilocus sequence typing, and resistance and virulence traits were investigated by PCR. Seven spa types were identified among MSSA (% of isolates): t127 (44.6), t605 (37.5), t002, t1784, t2066 (1.8), and 2 new ones: t10856 (10.7) and t10852 (1.8). Five distinct sequence types (ST) were detected (% of isolates): ST1 (46.4), ST126 (37.5), ST133 (10.7), ST5 (3.6), and a novel ST registered as ST2493 (1.8). Resistances were detected for streptomycin, chloramphenicol, and tetracycline. One strain contained the chloramphenicol resistance gene (fexA; included within transposon Tn558) and 3 strains contained the tetracycline resistance gene [tet(K)]. Methicillin-susceptible Staphylococcus aureus strains were susceptible to most of the antibiotics studied and lacked the virulence genes of Panton-Valentine leukocidin (lukF/S-PV), toxic shock syndrome toxin 1 (tst), exfoliative toxin A (eta), and exfoliative toxin B (etb), as well as the genes of the immune evasion cluster. Methicillin-susceptible Staphylococcus aureus isolates were detected in a relatively low proportion of cows with mastitis (6.2%) and recovered isolates presented high diversity of genetic lineages, with CC1 and CC126 the predominant clonal complexes, and CC133 also being detected. Larger epidemiological studies with molecular characterization of isolates are required to deepen the knowledge on the circulating genetic lineages among the cow population with mastitis. © 2013 American Dairy Science Association.

Perfil molecular e pesquisa de beta-lactamases de espectro ampliado de isolados de Klebsiella pneumoniase em rebanhos bovinos leiteiros no Estado de São Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Impacto da mastite subclínica na reprodução de vacas leiteiras

Pós-graduação em Zootecnia - FMVZ

Avaliação da eficácia do ácido lático frente ao iodo na anti-sepsia dos tetos após a ordenha na prevenção da mastite bovina

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Research of Klebsiella pneumoniae in dairy herds

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Investigation of biofilm production and icaA and icaD genes in Staphylococcus aureus isolated from heifers and cows with mastitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Intramammary coinfection by vaccinia virus and staphylococcus aureus in a bovine vaccinia outbreak

Introduction: Bovine vaccinia virus (VACV) is a well-known zoonotic agent related to exanthemous lesions in skin and mucous membranes of dairy cattle and humans, characterized by the formation of vesicles, pustules and ulcers. Mastitis is one of the most common infectious diseases of dairy herds. Bovine mammary infections are caused mainly by bacterial microorganisms, especially staphylococci. To the best of our knowledge, intramammary coinfection with VACV and Staphylococcus aureus in cows has not been reported previously. Case presentation: During an outbreak of exanthematic bovine VACV infection with animals showing vesicles, pustules and haemorrhagic ulcers on the teats, milk samples were collected for mastitis detection. Conclusion: The present report describes a case of intramammary coinfection by VACV and S. aureus in a bovine VACV outbreak.

The effect of phospholipase A2 from Crotalus durissus collilineatus on Leishmania (Leishmania) amazonensis infection

In this study, the effect of phospholipase A2 (PLA2) derived from Crotalus durissus collilineatus was evaluated in vitro and in vivo on experimental cutaneous leishmaniasis. The promastigote and amastigote forms treated with PLA2 presented increased growth rate. In vivo studies showed that PLA2-treated Leishmania (Leishmania) amazonensis promastigotes increased the size of lesions in BALB/c mice, and histopathological analysis showed numerous necrotic regions presenting a higher density of polymorphonuclear, mononuclear, and amastigote cells. Additionally, infected macrophages treated with PLA2 were able to generate prostaglandin E2 (PGE2). Cytokine quantification showed that the supernatant from infected macrophages presented moderate and high amounts of IL-2 and IL-10, respectively. However, in PLA2-treated infected macrophages, suppression of IL-2 levels occurred, but not of IL-10 levels. Observation also revealed that both the supernatant and lysate of L. (L.) amazonensis promastigotes exhibited PLA2 activity, which, in the presence of dexamethasone, showed no reduction in their activities; while glucocorticoid maintained the ability of promastigote forms to infect macrophages, which presented values similar to controls. In conclusion, the results indicate that PLA2 may be a progression factor for cutaneous leishmaniasis, since the PLA2 effect suppressed IL-2 levels and generated PGE2, an inflammatory lipid mediator.

Digenetic trematodes infection in a Bothrops moojeni (Viperidae) population from a fauna rescue in Porto Primavera, São Paulo State

No presente estudo é descrita a infecção por trematódeos digenéticos parasitas da cavidade oral e esôfago em uma população de serpentes Bothrops moojeni provenientes de resgate de fauna em Porto Primavera, Estado de São Paulo. Foi observada prevalência de infecção de 68%. O grau de infecção (número de trematódeos por serpente) variou de 2 a 51 helmintos. Os trematódeos encontrados foram Ophisthogonimus spp. e Sticholecitha serpentis. A alta prevalência de infecção foi associada com a drástica alteração ambiental e o estresse multi-fatorial aos quais os animais foram submetidos, que poderiam ter favorecido o ciclo dos parasitas.

Preferential infection sites of Cysticercus bovis in cattle experimentally infected with Taenia saginata eggs

The preferential sites of infection of Cysticercus bovis were evaluated in the skeletal muscle and entrails of 25 cattle that were experimentally infected with Taenia saginata (2 x 10(4) eggs). Two other animals were not inoculated (control). Ninety days after inoculation, all the cattle were euthanized. The carcasses were deboned and dissected into 26 anatomical sections (masseter muscles, brain, tongue, esophagus, heart, diaphragm, lungs, liver, kidneys, spleen, top sirloin butt, bottom sirloin butt, outside round, top (inside) round, transversus abdominus, top sirloin cap, strip loin, full tenderloin, eye of round, knuckle, shoulder clod, foreshank, shank, chuck, back ribs, and tail muscles). The dissected tissues were sliced into 5 mm sections. From the 25 cattle, 9258 C. bovis (cysticerci) were recovered; 75.02% (6946) of these were recovered from skeletal muscles and 24.98% (2312) from the entrails. A high parasitism level was found in the shoulder clod (12.55%), heart (11.02%), liver (9.48%), masseter muscles (8.51%), chuck (8.25%), strip loin and full tenderloin (7.26%), knuckle (6.63%), and back ribs (5.53%), totaling 69.23% (5738) of all of the detected cysticerci. on the other hand, there was a low C. bovis parasitism level in the brain, spleen, tail muscles, kidneys, esophagus, and diaphragm, representing just 3.9% of the total number of cysticerci. Given these results, we conclude that specific skeletal musculature regions, such as the shoulder blade, chuck, strip loin and full tenderloin, knuckle, back ribs and top round, which are not officially examined in many countries, are effective sites to efficiently screen C. bovis infection. To date, these regions have not been considered as preferential sites of C. bovis infection. Based on our work, however, these regions deserve greater attention from health inspectors because they contained a greater number of Cysticercus than the other regions of carcasses that are parasitized by T. saginata larvae. (c) 2010 Elsevier Ltd. All rights reserved.

Toxoplasma gondii: Infection natural congenital in cattle and an experimental inoculation of gestating cows with oocysts

Two studies, of a natural infection and an experimental infection, were performed in order to study congenital transmission of Toxoplasma gondii in cattle. In the first study, 50 fetuses were harvested from gestating cows that were eutanasied at a municipal slaughterhouse in Jaboticabal, São Paulo state, Brazil. In the second study, 11 gestating cows were divided into four groups for inoculation with T. gondii: GI consisted of three cows inoculated with 1.0 x 10(5) oocysts during their first trimester of gestation; GII consisted of three cows inoculated with 1.0 x 10(5) oocysts during their second trimester of gestation; GIII consisted of three cows inoculated with 1.0 x 10(5) oocysts during their last trimester of gestation; and GIV consisted of two control cows, one during its first and the other during its second trimester of gestation. In both studies, the presence of T. gondii was confirmed both indirectly by immunofluorescence assay (IFAT). In the natural infection experiment, 18% (9/50) of the gestating cows were confirmed to have specific antibodies (IFAT - 1:64) against T. gondii. The bioassay was able to diagnose the presence of T. gondii in the tissue samples from three calves. In the second experiment, the nine cows from groups I, II and III presented with specific antibodies (IFAT) against T. gondii. In contrast, T. gondii could not be detected by IFAT, histopathological examination or the bioassay in any of the nine calves born to cows experimentally infected with T. gondii oocysts. Based on the results from both studies, we conclude that congenital infection of T. gondii in cattle, while infrequent, does occur naturally. The pathogenicity of the strain of T. gondii may influence the likelihood of this route of transmission. (C) 2010 Elsevier B.V. All rights reserved.

Acute phase proteins: a potential approach for diagnosing chronic infection by Trypanosoma vivax

O objetivo do presente estudo foi verificar possíveis alterações nas proteínas de fase aguda em ovinos infectados experimentalmente com Trypanosoma vivax. Para tanto, foram utilizados oito ovinos machos, sendo quatro usados como controle e quatro infectados com 10(5) tripomastigotas de T. vivax. Colheram-se amostras de sangue em dois tempos antes da infecção e, posteriormente, aos 5, 7, 9, 11, 13, 15, 20, 30, 45, 60, 75, 90, 105 e 120 dias após a infecção (dpi); após centrifugação e aliquotização das amostras. As proteínas de fase aguda foram separadas por eletroforese em gel de acrilamida, contendo dodecil sulfato de sódio, e suas concentrações foram determinadas através de densitometria computadorizada. A dosagem de proteína total foi realizada pelo método colorimétrico do biureto. A contagem dos tripanossomas foi realizada diariamente, utilizando-se uma alíquota de 5 µL de sangue disperso em lâmina de microscopia, sob lamínula de 22 × 22 mm, contando-se os parasitos em 100 campos microscópicos, com objetiva de 40×, multiplicados pelo fator de correção do microscópio, e o resultado expresso em parasitos por mL de sangue. Para a análise estatística, empregou-se o teste de Wilcoxon a 5% de probabilidade. Foi observada a diminuição de diversas proteínas de fase aguda e aumento de antitripsina e transferrina que podem ser utilizadas para auxiliar no diagnóstico da infecção por T. vivax, principalmente na fase crônica da infecção.