Resistance to antimicrobials and biofilm formation in Staphylococcus spp. isolated from bovine mastitis in the Northeast of Brazil.

Mastitis is the principal disease affecting dairy herds worldwide. The aim of the present study was to characterize phenotypic and genotypic features associated with resistance to antimicrobials in Staphylococcus spp. isolated from 2064 milk samples of 525 lactating cows in the Northeast of Brazil. Of the 218 isolates analyzed, 57.8% were characterized as Staphylococcus aureus, 28% as coagulase-positive staphylococci other than S. aureus (oCPS), and 14.2% as coagulase-negative staphylococci (CNS). The test for susceptibility to antimicrobials showed amoxicillin (32.6%) to be the less effective drug in vitro, and the multi-drug resistance (MDR) rate for beta-lactams varied from 0 to 0.75. The genotypic characterization showed that 93.1% of the samples were tested positive for the blaZ gene, while none amplified mecA. The antibiotic efflux mechanism was observed in 0.9% of isolates. The biofilm formation was found in 3.7 and 96.3% of samples, respectively, on Congo red agar and on the microplate adhesion test, while the icaD gene was present in 92.2% of Staphylococcus spp. The high frequency of blaZ gene observed in this study was associated with the resistance of most Staphylococcus spp. to one or more of the beta-lactams tested, which are routinely used in Brazilian herds for mastitis treatment. The biofilm formation was also detected in the isolates analyzed being an important characteristic for pathogenicity and antimicrobial resistance of bacteria.

Toxoplasma gondii in invasive animals on the Island of Fernando de Noronha in Brazil: Molecular characterization and mouse virulence studies of new genotypes.

This study aimed to genetically characterize and to determine virulence from Toxoplasma gondii samples from invasive animals in the Island of Fernando de Noronha, Brazil. Blood samples were collected from 21 tegu-lizard (Salvator merianae), 12 rock-cavies (Kerodon rupestris) and 154 black-rats (Rattus rattus) from the Island and MAT (cutoff 1:25) detected anti-T. gondii antibodies in 0% of the tegus (0/21); 58.3% of the rock-cavies (7/12) and 22.7% of rats (35/154). Tissue samples (brain, heart, liver and lung) from positive animals in MAT were collected for molecular analysis and for bioassay in Swiss Webster mice. After observation period, mice were euthanized, and serological detection and tissue cyst search in the brain were performed. The brain of positive animals for serological detection or tissue cyst search was cultured in MARC-145 cells for maintenance of the T. gondii isolate. No isolate was obtained from rock cavies. Nine isolates were obtained by bioassay of 35 seropositive black rats. DNA samples were extracted from rat tissues and from parasite isolates in cell culture, and genotyped using 10 PCR-RFLP markers. ToxoDB genotypes #78 (1) from rat tissue and #146 (4), #163 (2), #260 (2) and #291 (1) from cell culture were detected. Markers of genes ROP18 and ROP5 were analyzed and in vivo virulence test was conducted in mice. Analysis revealed two allele combinations, 3/1 and 3/3, indicating non-lethal T. gondii strains, which is supported by mouse virulence test.

High prevalence of toxoplasmosis in free-range chicken of the Fernando de Noronha Archipelago, Brazil.

The present study aimed to determine the prevalence of toxoplasmosis in free-range chickens of the Fernando de Noronha Archipelago, Brazil and to identify risk factors associated with Toxoplasma gondii infection. Blood samples were collected from all the adult chickens raised in the Archipelago and screened by Indirect Fluorescent Antibody Test (430 samples, in total). Univariate analysis (Chi-square) and logistic regression were used to investigate the relationship between various variables possibly predictive of an increased likelihood of T. gondii infection. The overall prevalence of T. gondii infection in chickens of the Fernando de Noronha Archipelago was 88.4% (380/430; 84.6%-91.0%; 95% CI), ranging from 57.1% to 100.0% among the studied properties. The risk factors associated with T. gondii infection were the number of domestic cats in the properties (p=0.022), the presence of feral cats (p=0.006) and the presence of an open water source (p=0.046). Domestic and feral cats should be prevented from accessing the water and food supplied to chickens.

Spatial characterization of Leptospira spp. infection in equids from the Brejo Paraibano micro-region in Brazil.

The present study, the first to spatially characterize Leptospira spp. infection among equids in the Brejo Paraibano micro-region of the Paraiba state in the northeast of Brazil, investigated 257 animals in 26 farms properties. Serum samples from 204 horses, 46 mules and seven donkeys were serologically diagnosed using the microscopic agglutination test (MAT). The distribution of Leptospira spp. was studied by employing specific antigens from 24 different Leptospira serovars. All farms were georeferenced and their distribution visualised on a map of the Brejo Paraibano micro-region. In addition, rainfall data were obtained from the same year, in which the sampling was performed. Among the 20 farms found to harbour animals with leptospirosis, 14 (70%) exhibited low prevalence, five (25%) medium prevalence and one (5%), high prevalence. Certain areas had a higher density of infected farms and required intervention to control the infection. Many serovars were widely distributed, while others were more common in particular areas. There was no significant association between the prevalence of Leptospira spp. infection and rainfall.

Occurrence of anti-Neospora caninum antibodies in cattle in the dairy farming region of the state of Piauí, Brazil

Abstract This study focused on the detection of anti-Neospora caninum IgG antibodies in cows in the dairy farming region of the state of Piauí, Brazil. To this end, serum samples were collected from 255 dairy cows on 17 farms located in the dairy farming region of the municipality of Parnaíba, Piauí, Brazil. The indirect fluorescence antibody test (IFAT) was employed to detect anti-N. caninum IgG antibodies, using anti-bovine IgG (Sigma®) conjugated to fluorescein isothiocyanate (FITC) and a cutoff point of 1:200. Of the 255 samples analyzed, 69 (27.06%) were positive for anti- N. caninum IgG antibodies, the relative frequency found by property was: 1 (20.00%), 2 (53.33%), 3 (46.66%), 4 (53.33%), 5 (26.66%), 6 (6.66%), 7 (6.66%), 8 (20.00%), 9 (26.66%), 10 (26.66%), 11 (20.00%), 12 (20.00%), 13 (46.66%), 14 (26.66%), 15 (26.66%), 16 (20.00%) and 17 (13.33%). with titers of 200 (15.94%), 400 (20.30%), 800 (24.63%), 1600 (23.18%) and 3200 (15.94%), being the highest frequency for the titer of 800. This study demonstrates for the first time that cows from dairy herds of Parnaíba municipality, state of Piauí, are exposed to N. caninum.

Resumo Objetivou-se estudar a ocorrência de anticorpos IgG anti- Neospora caninum em vacas, na bacia leiteira do estado do Piauí, Brasil. Foram obtidas amostras séricas de 255 vacas leiteiras provenientes de 17 propriedades localizadas na bacia leiteira do município de Parnaíba, Piauí, Brasil. Para a pesquisa de anticorpos IgG anti- N. caninum foi empregada a técnica de Reação de Imunofluorescência Indireta, utilizando-se conjugado anti-IgG bovine (Sigma®) conjugado ao isotiocianato de fluoresceína e ponto de corte 1:200. Das 255 amostras analisadas, 69 (27,06%) foram positivas para anticorpos IgG anti- N. caninum, a frequência relativa encontrada por propriedade foi de: 1 (20,00%), 2 (53,33%), 3 (46,66%), 4 (53,33%), 5 (26,66%), 6 (6,66%), 7 (6,66%), 8 (20,00%), 9 (26,66%) 10 (26,66%), 11 (20,00%), 12 (20,00%), 13 (46,66%), 14 (26,66%), 15 (26,66%), 16 (20,00%) e 17 (13,33%) com títulos de 200 (15,94%), 400 (20,30%), 800 (24,63%), 1600 (23,18%) e 3200 (15,94%), sendo a maior frequência para o título de 800. Este estudo demonstra pela primeira vez que, vacas de rebanhos leiteiros do município de Parnaíba, estado do Piauí, estão expostas ao N. caninum.

Isolation and genetic characterization of Toxoplasma gondii from free-ranging and captive birds and mammals in Pernambuco state, Brazil

Abstract Recent genetic population studies on Toxoplasma gondii in Brazil have shown large genetic variability. The objective of the present study was to isolate and genotypically characterize T. gondii from free-ranging and captive wild mammals and birds in Pernambuco state, Brazil. Fragments of heart, brain, skeletal muscle and diaphragm tissue from 71 birds and 34 mammals, which were either free-ranging or captive, were collected. Samples from 32 of these animals were subjected to bioassays in mice. Samples from the remaining 73 animals underwent biomolecular diagnosis, using PCR technique, targeting a repetitive DNA fragment of 529 bp in T. gondii. A non-virulent isolate (TgButstBrPE1) was obtained from a free-ranging striated heron (Butorides striata) and, based on primary samples, seven animals were found to be positive. The primary samples and the isolate obtained were subjected to PCR-RFLP using the markers SAG1, 53SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. ToxoDB-RFLP genotype /13 from the striated heron isolate and Type BrIII genotype from a captive otter ( Lontra longicaudis) (PS-TgLonloBrPE1) were obtained. The present study describes the first isolation and genotypic characterization of T. gondii in free-ranging striated heron, and the first genotypic characterization of T. gondii in a captive otter.

Resumo Recentes estudos genéticos nas populações deste parasita no Brasil têm mostrado grande variabilidade genética. O objetivo do presente estudo foi isolar e caracterizar genotipicamente T. gondii de aves e mamíferos de vida livre e de cativeiro no estado de Pernambuco, Brazil. Fragmentos de tecido do coração, cérebro, músculo esquelético e diafragma de 71 aves e 34 mamíferos de vida livre ou cativeiro foram colhidos. Amostras de 32 destes animais foram submetidas a bioensaios em camundongos. As amostras dos 73 animais restantes foram submetidas a diagnóstico biomolecular usando a técnica de PCR, tendo como alvo o fragmento repetitivo de 529 pb do DNA de T. gondii. Dentre os 32 bioensaios conduzidos, obteve-se um isolado não-virulento (TgButstBrPE1) de um socozinho (Butorides striata ) de vida livre, e dentre as amostras primárias, sete animais foram positivos. As amostras primárias e o isolado foram submetidos a PCR-RFLP usando os marcadores SAG1, 53SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico e CS3. Foram obtidos o genótipo ToxoDB-RFLP /13 do isolado do socozinho e o genótipo Type BrIII de uma lontra (Lontra longicaudis) de cativeiro (PS-TgLonloBrPE1). O presente estudo descreve o primeiro isolamento e caracterização genotípica de T. gondii em socozinho de vida livre, e a primeira caracterização genotípica de T. gondii em lontra em cativeiro.

Prevalência e intensidade média de infecção de ciatostomíneos (Cyathostominae: Strongylidae) do cólon ventral de equídeos provenientes do estado de Pernambuco (dados preliminares)

As sete espécies de ciatostomíneos mais prevalentes do cólon ventral em Equus caballusforam Cyathostomum tetracanthume Cylicostephanus minutus, ambas com prevalência de 85,7%, Cylicocyclus nassatus (71 ,4%) e prevalência de 75,1% paraas espécies Coronocyclus labiatus, Coronocyclus /abratus, Cylicocyclus leptostomus e Cylicostephanus calicatus. Em Equus asinus, C. minutus, C. nassatus e C. /eptostomus apresentaram prevalência de 100%.

Infecções micóticas e bacterianas em lesões cutâneas de cães parasitologicamente positivos para Leishmania chagasi

Este trabalho teve como objetivo verificar a ocorrência de infecção cutânea fúngica e bacteriana em cães parasitologicamentepositivos para leishmaniose visceral canina (LVC). Para o diagnóstico da LVC foram realizados raspados de pele íntegra e/oulesionada, punção de medula óssea e punção aspirativa de linfonodo poplíteo, utilizando-se 46 animais. Paralelamente foicoletado material para cultura fúngica e bacteriana. Obtevese 23,91% de positividade para LVC, sendo 90,90% dos casosdiagnosticados por punção de medula óssea, 45,45% por raspado de pele íntegra e 36,36% através de raspado de pelelesionada. Observouse que associadas às lesões de LVC, ocorreram concomitantemente infecções fúngicas (66,66%) ebacterianas (100%), predominantemente por Aspergillus sp. e Staphylococcus sp. respectivamente.

Plasmodium juxtanucleare (Versiani e Gomes, 1941) em galinhas ( Gallus gallus L., 1857) de criações rústicas no Estado de Pernambuco

O Plasmodium juxtanucleare é um dos agentes etiológicos da Malária Aviária, que se caracteriza clinicamente por anemia,tristeza, inapetência, incoordenação motora, prostração e morte. Este estudo teve como objetivo determinar a ocorrência desteparasito no Estado de Pernambuco. Foram analisados 35 esfregaços sangüíneos, corados pelo método de Giemsa, degalinhas de criações rústicas da Região Metropolitana do Recife. As formas eritrocitárias do parasito observadas foram:trofozoítos, esquizontes e gametócitos. As aves apresentaram baixa parasitem ia, caracterizando doença crônica e assintomática.Do total das amostras analisadas, 51,4% foram positivas. Descreve-se, pela primeira, vez a ocorrência do Plasmodiumjuxtanucleare no Estado de Pernambuco - Brasil.

Biopolímero produzido a partir da cana-de-áçucar para cicatrização cutânea

An extracelular polysaccharide was produced through microbiology, using the ZSP bacteria isolated in the Industrial Microbiological Laboratory of the Sugar cane Experimental Station at Carpina/UFRPE, Pernambuco, Brazil, presenting exceptional process capacity. The principal monosaccharides present in the ¹soluble fraction were glucose (87.6%), xylose (8.6%), mannose (0.8%), ribose (1.7%), galactose (0.1%), arabinose (0.4%) and the glucuronic acid (0.8%). Due to the high rate of injuries that occur with domestic animals and the search for simple, economical alternatives that would be capable of giving ideal conditions for the healing process, extensive testing was done with the biopolymer produced by sugar cane in animals that had cutaneous wounds, so as to evaluate the reepitelization process. It was observed from the testing results that there was better skin granulation, better infection control, and less healing time, which allowed the conclusion that the biopolymer contributed favorabably in the healing process, and could be used in cutaneous injuries.

Um polissacarídeo extracelular foi produzido por via microbiológica, através da bactéria ZSP isolada no Laboratório de Microbiologia Industrial da Estação Experimental de Cana-de-Açúcar do Carpina/UFRPE, apresentando excepcional capacidade de processo. Os principais monossacarídeos presentes na fração solúvel foram glicose (87,6%), xilose (8,6%), manose (0,8%), ribose (1,7%), galactose (0,1%), arabinose (0,4%) e o ácido glucurônico (0,8%). Devido ao alto índice de traumatismos que acomete os animais domésticos e a busca por alternativas simples, econômicas e capazes de proporcionar condições ideais para cicatrização, foram realizados os testes com o biopolímero produzido a partir da cana-de-açúcar em animais portadores de feridas cutâneas, a fim de avaliar a reepitelização. Observou-se o aumento do tecido de granulação, controle da infecção e diminuição do tempo de cicatrização, permitindo concluir que o biopolímero contribui para o processo cicatricial, podendo ser utilizado em feridas cutâneas.