Salmonella enterica in Invasive Lizard from Fernando de Noronha Archipelago: Serotyping, Antimicrobial Resistance and Molecular Epidemiology.

Salmonella infection can pose serious health issues, especially to children, elders or immunosuppressed humans. Wild populations of reptiles can reach Salmonella prevalence of up to 100% and the direct or indirect transmission from reptiles to humans have been extensively reported. Fernando de Noronha (FN) is an inhabited oceanic archipelago in the northeast coast of Brazil, with an economy based on tourism. The tegu (Salvator merianae) is the largest lizard native to South America and was introduced to the archipelago in the early 20th century. This study determines the prevalence, serotypes, antimicrobial resistance, and molecular epidemiology of Salmonella enterica in the tegu population from FN archipelago. Results show that S. enterica is widely distributed in the FN tegu population, with 43.8% prevalence. The bacteria were isolated from 70.5% of the sampled sites and a total of 15 serotypes were detected in 98 S. enterica isolates. Strains were further classified into 31 genotypes. Recaptured animals presented distinct genotypes in each season, demonstrating a seasonal strain turnover. Most S. enterica isolates from FN tegus presented low antimicrobial resistance. This is possibly due to geographical isolation of the island population, hampering contact with strains from livestock from the continent, where antimicrobial resistance is common.

Feline Sporothrix spp. detection using cell blocks from brushings and fine-needle aspirates: Performance and comparisons with culture and histopathology.

BACKGROUND: Sporotrichosis is an emerging zoonotic mycosis that presents as a cutaneous lymphatic or disseminated disease, caused by fungi from the Sporothrix schenkii (S schenkii) clinical clade. Its importance is growing, primarily due to an outbreak that occurred in Brazil, affecting mainly cats and people. OBJECTIVES: In Brazil, an S schenkii diagnosis is often made using cultures, which allows genus identification and sufficient growth to perform molecular biology testing. Despite its advantages, fungal cultures are slow to develop and can delay public health measures, highlighting the importance of developing additional diagnostics techniques. METHODS: Cell block cytology (CBLC) is an older method that regained importance after liquid-based cytology (LBC) was introduced, and it has been previously and successfully applied to veterinary diagnostics. We aimed to standardize and compare CBLC from cervical brush exfoliation of open wounds and fine-needle aspirates with culture and immunohistochemistry of skin biopsies for sporotrichosis in cats, as a novel method. RESULTS: For this purpose, we selected 40 cats with skin lesions suspected of having sporotrichosis in Guarulhos city, São Paulo state, Brazil. We achieved 97.5% and 95% positivity using CBLC and culture, respectively, and 100% of feline skin biopsies were positive for Sporothrix spp on histopathology/immunohistochemistry. CONCLUSIONS: Cell block cytology is an efficient and rapid tool to diagnose sporotrichosis in cats, particularly during epidemics.

Prevalence and molecular features of ESBL/pAmpC-producing Enterobacteriaceae in healthy and diseased companion animals in Brazil.

Extended-spectrum beta-lactamase (ESBL)- and plasmid-mediated AmpC (pAmpC)-carrying Enterobacteriaceae have widely disseminated in human, animal and environmental reservoirs. Pets have been recognized as a source of ESBL/pAmpC worldwide, and are possibly also a source of human contamination. The aim of this study was to document to what extent cats and dogs may act as a driving force in the spread of ESBLs and pAmpCs in Brazil. A total of 113 healthy stray cats and dogs and 74 sick pets were sampled, and extended-spectrum cephalosporin-resistant Enterobacteriaceae (ESC-R) were detected in 28/113 (24.8%) and 8/74 (10.8%) tested animals, respectively. Different Enterobacteriaceae isolates (mostly E. coli), a large number of E. coli clones (with ST90, ST457, ST973 and ST2541 being predominant), and several ESBL/pAmpC genes and plasmids were characterized, highlighting the ability of stray and pet cats and dogs to further spread a wide range of ESC-resistance determinants. The ESBL phenotype was due to the blaCTX-M-2 and blaCTX-M-8 genes, as found in human epidemiology in Brazil, but blaCTX-M-9 and blaCTX-M-15 were also identified. The pAmpC phenotype was systematically due to the presence of the blaCMY-2 gene, mostly carried by IncI1 ST12 plasmids. Our results showed that pets can be considered a significant reservoir of multidrug-resistant bacteria in Brazil. This is especially true for healthy stray dogs that displayed the highest prevalence (24.8%) of ESBLs/pAmpC resistance determinants, which can then be further spread both to the environment and to other animals or humans by contact.

An Atypical Enteropathogenic Escherichia coli Isolate with Possible Human and Domestic Animal Origin from a Free-Living Lear's Macaw (Anodorhynchus leari).

An atypical enteropathogenic Escherichia coli was isolated from an asymptomatic nestling of the endangered Lear's Macaw (Anodorhynchus leari). Phylogenetically, it was identical to bovine and human strains and highly similar to other human and domestic animal isolates. We discuss potential routes of transmission and risks to conservation of this species.

High dilutions of antimony modulate cytokines production and macrophage - Leishmania (L.) amazonensis interaction in vitro.

BACKGROUND: In previous results mice treated with high dilutions of antimony presented reduction of monocyte migration to the site of infection with increase in B lymphocytes population in the local lymph node. AIMS: To know the mechanisms involved, a series of in vitro studies was done, using co-cultures of macrophages (RAW 264.7) and Leishmania (L.) amazonensis treated with different dilutions of antimony (Antimonium crudum or AC), in different times. METHODOLOGY: Spreading, phagocytosis, the oxidative activity of macrophages, the viability of free promastigotes and the cytokines/chemokines concentration in the supernatant were evaluated. The assays were performed in quadruplicate. RESULTS: Cells treated with AC 30cH (10-58M) and AC 200cH (10-398M) presented a temporary reduction of the spreading after 02h of incubation, followed by increase after 48h, being the most significant increase observed after the AC 200cH treatment. However, the percentage of internalized parasites at 48, 96 and 120h of incubation was also higher in cells treated with AC 200cH. It is suggested that the AC 200cH improves the ability of phagocytes to internalize the parasites, but not to digest them. The cytokines-chemokines panel corroborated these results. Both dilutions potentiated the parasite-induced reduction of cytokines production, especially IL-6, IL 12 p40 and γ-IFN, after 48h of incubation. In addition, the production of MIP-1 beta (CCL4), a chemokine involved in chronic inflammation, was also reduced after 120h. A specific effect of AC 30cH was seen by the inhibition of two peaks of CCL2 (MCP-1) observed in infected macrophages, at 24 and 120h. Since this cytokine is an important chemokine for monocytes, it explains the results obtained formerly in vivo. The morphology of macrophages after acridine orange staining revealed that the treatment with AC 30cH reduced substantially the acid vacuoles in the cytoplasm, indicating a certain inability of these cells to digest the parasites. On the other hand, a large peak of VEGF-A, associated with increase of internalized parasites was observed after 120h of treatment with AC 200cH, which could be associated to the regulation of the chronic inflammation events by M1-M2 polarization. There was no statistical difference among groups regarding the production of TNF, NO and H2O2, showing that the drugs do not alter macrophage cytotoxic activity. A clear quantitative and qualitative variation of the modulatory effects of AC 30cH and 200cH was seen, in function of time. CONCLUSIONS: Both dilutions were able to potentiate the decrease of most of cytokines and chemokines induced by the parasite infection in vitro, which explains the clinical improvement seen previously in vivo, however, the mechanisms involved and the epidemiological significance of these findings are still under discussion.

Detection of bacteria and fungi and assessment of the molecular aspects and resistance of Escherichia coli isolated from confiscated passerines intended for reintroduction programs.

Many native bird species are currently considered rare in Brazil because they have been indiscriminately collected by animal traffickers and commercialized, leading to dwindling numbers in their natural habitats. Confiscated animals are at times destined for reintroduction programs that must ensure these animals do not pose a risk to native populations. Healthy or sick wild passerines may carry a great diversity of microorganisms. Therefore, knowledge of the sanitary status of confiscated animals destined for reintroduction is critical to assess whether these animals act as microorganism carriers and to investigate the epidemiology of transmissible diseases, a crucial aspect for animal and human health preservation. This study examined the occurrence of aerobic and facultative anaerobic bacteria and fungi in cloacal swabs collected from wild confiscated passerines intended for reintroduction programs. In vitro susceptibility tests of the most frequent isolates as well as studies of the molecular aspects of Escherichia coli isolates were also performed. There was microorganism growth in 62.5% of 253 samples. The microorganisms that were most frequently isolated were Staphylococcus spp. (15.0%), Micrococcus spp. (11.5%), E. coli (10.7%) and Klebsiella spp. (10.7%). Fifteen bacteria genera and seven fungi genera were isolated. Multidrug-resistance to antimicrobials was observed in Staphylococcus spp., Micrococcus spp., E. coli and Klebsiella spp. isolates. The high occurrence of Enterobacteria observed is possibly related to the sanitary conditions in which confiscated animals are usually kept. One E. coli sample (out of 27 isolates) was positive for the S-fimbrial adhesion encoding gene (sfa). Considering the low occurrence of genes that encode virulence factors, confiscated passerines may represent a low risk for the potential transmission of EPEC, APEC, UPEC and NMEC isolates to other animals or humans. The potential risk of intra- or inter-specific transmission of multidrug-resistant isolates and the introduction of these microorganisms into the environment must be considered, although there are still therapeutic alternatives for treatment of these animals among the antimicrobials which were tested. The stress and poor hygiene conditions imposed on animals during trafficking may have caused their contamination by multidrug-resistant agents transmitted by humans or by the precarious environment to which they were subjected. Risks related to the dissemination of Salmonella spp., Cryptococcus spp. and Candida spp. are low when reintroduction programs are considered.

Microbiota bacteriana e citologia da região traqueobrônquica de bezerros no período neonatal / Bacterial pathogens and cytology or tracheobronchial region in calves during neonatal period

As broncopneumonias são afecções importantes na pecuária mundial, representando uma das principais causas de mortalidade de bezerros nos primeiros meses de vida. As medidas preventivas e terapêuticas adotadas geralmente são baseadas em resultados de estudos internacionais, não se conhecendo as bactérias implicadas nos quadros pneumônicos em animais criados no Brasil. Aliado a isso, no primeiro mês de vida, os bezerros demonstram imaturidade do sistema imune, o que tem sido pouco estudado em quadros pneumônicos. Desta maneira, objetivou-se estudar as broncopneumonias em bezerros neonatos, identificando bactérias do trato respiratório posterior de bezerros sadios e com pneumonias naturalmente adquiridas, bem como analisar citologicamente a resposta pulmonar frente a estes patógenos. Para isso amostras de lavado do trato respiratório foram colhidas por traqueocentese durante o primeiro mês de vida dos animais. Verificou-se que não houve diferença na microbiota traqueobrônquica de bezerros sadios em relação aos doentes, discordando dos relatos da literatura internacional, sendo constituída principalmente por: Staphylococcus sp., Bacillus sp., Streptococcus sp., Pseudomonas aeruginosa e enterobactérias, permitindo inferir que as medidas profiláticas e terapêuticas adotadas internacionalmente possam não ser tão efetivas para as criações brasileiras. Observou-se também que bezerros neonatos têm uma proporção aproximada de 1:1 de macrófagos e neutrófilos na região traqueobrônquica quando saudáveis, atingindo uma relação aproximada de 1:3 durante os quadros de broncopneumonias, sendo estes perfis provavelmente característicos da idade, período conhecido pela imaturidade do sistema imune e agravado por fatores de manejo que favoreçam uma maior inalação de agentes bacterianos.

Bronchopneumonia is important for world livestock production and one of the major causes of calf mortality during the first months of life. The preventive and therapeutic measures adopted for the disease in calves in Brazil are usually based on the results of international studies; however there is not enough known which bacteria are implicated. In the first month of life calves show immaturity of their immune system, what has been little studied in regard to pneumonia. The present investigation sought to study neonate bronchopneumonia in calves, to identify which bacteria are present in the respiratory tract of healthy and, with naturally acquired pneumonia calves, to analyze the pulmonary cytological response against pathogens. For this purpose, samples of the respiratory tract were collected by tracheocenthesis. It was noted that the microflora of the tracheobronchial region of healthy and diseased calves was the same, but they were different from the one reported by international papers. The flora was constituted mainly by Staphylococcus sp., Bacillus sp., Streptococcus sp., Pseudomonas aeruginosa and enterobacteriae, allowing to infer that the prophylactic and therapeutic measures adopted internationally may not be as effective for the Brazilian condition. It was also found that newborn calves have an approximate ratio of 1:1 of macrophages and neutrophils in the tracheobronchial region when they were healthy, reaching a ratio of approximately 1:3 of macrophages and neutrophils when they were with bronchopneumonia. Probably, these profiles are characteristic of the age, a period when exists immaturity of the immune system and influenced by management factors which lead to greater inhalation of bacterial agents.

Microbiota bacteriana e citologia da região traqueobrônquica de bezerros no período neonatal / Bacterial pathogens and cytology or tracheobronchial region in calves during neonatal period

As broncopneumonias são afecções importantes na pecuária mundial, representando uma das principais causas de mortalidade de bezerros nos primeiros meses de vida. As medidas preventivas e terapêuticas adotadas geralmente são baseadas em resultados de estudos internacionais, não se conhecendo as bactérias implicadas nos quadros pneumônicos em animais criados no Brasil. Aliado a isso, no primeiro mês de vida, os bezerros demonstram imaturidade do sistema imune, o que tem sido pouco estudado em quadros pneumônicos. Desta maneira, objetivou-se estudar as broncopneumonias em bezerros neonatos, identificando bactérias do trato respiratório posterior de bezerros sadios e com pneumonias naturalmente adquiridas, bem como analisar citologicamente a resposta pulmonar frente a estes patógenos. Para isso amostras de lavado do trato respiratório foram colhidas por traqueocentese durante o primeiro mês de vida dos animais. Verificou-se que não houve diferença na microbiota traqueobrônquica de bezerros sadios em relação aos doentes, discordando dos relatos da literatura internacional, sendo constituída principalmente por: Staphylococcus sp., Bacillus sp., Streptococcus sp., Pseudomonas aeruginosa e enterobactérias, permitindo inferir que as medidas profiláticas e terapêuticas adotadas internacionalmente possam não ser tão efetivas para as criações brasileiras. Observou-se também que bezerros neonatos têm uma proporção aproximada de 1:1 de macrófagos e neutrófilos na região traqueobrônquica quando saudáveis, atingindo uma relação aproximada de 1:3 durante os quadros de broncopneumonias, sendo estes perfis provavelmente característicos da idade, período conhecido pela imaturidade do sistema imune e agravado por fatores de manejo que favoreçam uma maior inalação de agentes bacterianos.(AU)

Bronchopneumonia is important for world livestock production and one of the major causes of calf mortality during the first months of life. The preventive and therapeutic measures adopted for the disease in calves in Brazil are usually based on the results of international studies; however there is not enough known which bacteria are implicated. In the first month of life calves show immaturity of their immune system, what has been little studied in regard to pneumonia. The present investigation sought to study neonate bronchopneumonia in calves, to identify which bacteria are present in the respiratory tract of healthy and, with naturally acquired pneumonia calves, to analyze the pulmonary cytological response against pathogens. For this purpose, samples of the respiratory tract were collected by tracheocenthesis. It was noted that the microflora of the tracheobronchial region of healthy and diseased calves was the same, but they were different from the one reported by international papers. The flora was constituted mainly by Staphylococcus sp., Bacillus sp., Streptococcus sp., Pseudomonas aeruginosa and enterobacteriae, allowing to infer that the prophylactic and therapeutic measures adopted internationally may not be as effective for the Brazilian condition. It was also found that newborn calves have an approximate ratio of 1:1 of macrophages and neutrophils in the tracheobronchial region when they were healthy, reaching a ratio of approximately 1:3 of macrophages and neutrophils when they were with bronchopneumonia. Probably, these profiles are characteristic of the age, a period when exists immaturity of the immune system and influenced by management factors which lead to greater inhalation of bacterial agents.(AU)

Proteinase and phospholipase activities and development at different temperatures of yeasts isolated from bovine milk.

The presence of yeasts in milk may cause physical and chemical changes limiting the durability and compromising the quality of the product. Moreover, milk and dairy products contaminated by yeasts may be a potential means of transmission of these microorganisms to man and animals causing several kinds of infections. This study aimed to determine whether different species of yeasts isolated from bovine raw milk had the ability to develop at 37°C and/or under refrigeration temperature. Proteinase and phospholipase activities resulting from these yeasts were also monitored at different temperatures. Five genera of yeasts (Aureobasidium sp., Candida spp., Geotrichum spp., Trichosporon spp. and Rhodotorula spp.) isolated from bovine raw milk samples were evaluated. All strains showed one or a combination of characteristics: growth at 37°C (99·09% of the strains), psychrotrophic behaviour (50·9%), proteinase production (16·81% of the strains at 37°C and 4·09% under refrigeration) and phospholipase production (36·36% of the isolates at 37°C and 10·9% under refrigeration), and all these factors may compromise the quality of the product. Proteinase production was similar for strains incubated at 37°C (16·81% of the isolates) and room temperature (17·27%) but there was less amount of phospholipase-producing strains at room temperature (15·45% of the isolates were positive) when compared with incubation at 37°C (36·36%). Enzymes production at 37°C by yeasts isolated from milk confirmed their pathogenic potential. The refrigeration temperature was found to be most efficient to inhibit enzymes production and consequently ensure better quality of milk. The viability of yeasts and the activity of their enzymes at different temperatures are worrying because this can compromise the quality of dairy products at all stages of production and/or storage, and represent a risk to the consumer.

Correlation between mastitis occurrence and the count of microorganisms in bulk raw milk of bovine dairy herds in four selective culture media.

Milk is the normal secretion of the mammary gland, practically free of colostrum and obtained by the complete milking of one or more healthy animals. Mastitis is an inflammatory process of the mammary gland and it may cause alterations in the milk. The present work aimed to verify whether it is possible, by means of the counts of microorganism in the bulk raw milk in four selective culture media, to establish a correlation with the occurrence of mastitis and therefore, to monitor this disease in bovine dairy herds. The following selective culture media were used: KF Streptococcus Agar, Edwards Agar, Baird-Parker Agar, Blood Agar plus potassium tellurite. Spearman's correlation coefficient was calculated in order to compare the occurrence of mastitis (percentage) in each herd with respective selective culture media counts of microorganisms in bulk raw milk. Thirty-six possibilities were analysed (Tamis and CMT-positive rates were compared with the log-transformed count in four selective culture media) and there was a negative correlation between Tamis 3 and the Baird-Parker Agar plate count. The total results of microbiological tests showed that there were three correlations of the counts in selective culture media. Fifty-two possibilities were analysed and there was a negative correlation between no-bacterial-growth mastitis rates and log10 of KF Streptoccocus Agar plate count and there were two positive correlations between coagulase-positive staphylococci and log10 of Baird-Parker Agar plate count and Blood Agar plus potassium tellurite plate count.

Perfil celular e microbiológico do leite de ovelhas Santa Inês no período lactante e pós-desmame / Cellular and microbiological profile of Santa Ines ewes in the lactation and the post-weaning period

The aim was to evaluate and compare the microbiological and cellular profile of the milk of Santa Ines ewes during the lactation period and the active involution. Milk samples were analyzed from 12 ewes during these distinct periods. Clinical examination of the mammary gland, somatic cell count (SCC), California Mastitis Test (CMT), bacteriologic screening and sensibility of the pathogens in vitro were performed. Most alterations were observed in the active involution period. SCC and CMT were higher in this same period. Besides this, a high persistency of infection occurred. The active involution period did not show high susceptibility. Coagulase-negative staphylococci were the only isolated bacteria. A high antimicrobial sensibility of these pathogens was also encountered.

O objetivo deste trabalho foi avaliar o perfil microbiológico e celular do leite no período lactante e de involução ativa de ovelhas da raça Santa Inês. Foram avaliadas amostras lácteas de 12 ovelhas durante estes distintos períodos. Realizou-se o exame físico da mama, sendo as amostras lácteas submetidas à contagem de células somáticas (CCS), ao California Mastitis Test (CMT), ao exame microbiológico e aos testes de sensibilidade in vitro dos patógenos encontrados. Foram observados maiores escores do exame físico, CCS, CMT durante o período de involução ativa, além de uma alta persistência da infecção durante estes períodos. O período de involução ativa não se mostrou como um momento de alta susceptilidade. Os estafilococos coagulase negativa representaram o único gênero isolado das glândulas infectadas. Uma alta sensibilidade dos agentes etiológicos envolvidos frente aos diferentes antimicrobianos in vitro foi também observada.

Relationship between occurrence of mastitis pathogens in dairy cattle herds and raw-milk indicators of hygienic-sanitary quality.

Mastitis is an inflammation of the mammary glands and in most cases it is caused by the presence of microorganisms. High mastitis rates in dairy cattle herds can cause an increase in total microorganism counts of bulk tank milk. The present paper was aimed at verifying whether the occurrence of mastitis in dairy cattle herds is reflected in raw-milk indicators of hygienic-sanitary quality. To observe the correlation among the analysed variables, we performed a logarithmical transformation (log10) of different indicator counts of raw milk and compared them with the occurrence of mastitis in dairy cattle herds. Few correlations were observed among mastitis cases in dairy cattle herds and the raw-milk indicators of hygienic-sanitary quality. We observed a negative correlation between the log10 of mesophilic aerobic plate counts and psychotropic aerobic plate counts when compared with the occurrence of no bacterial growth. The log10 of thermophilic aerobic plate counts and yeasts and mould aerobic plate counts presented a positive correlation with the cases of infectious mastitis and mastitis caused by Staphylococcus spp.

Tratamento homeopatico de melanoma maligno em cadela / Homeopathic treatment of malignant melanoma in a dog

O melanoma maligno e o tumor oral mais comumente diagnosticado em caes. A eficiencia dos tratamentos de melanoma maligno pela excisao cirurgica, radioterapia ou quimioterapia e muito pequena e o prognostico ruim. O objetivo do... (AU)