Maria Helena Cosendey de Aquino

Campylobacter jejuni/coli : Methodology of isolation and possible interfering factors in primary culture

The prevalence of Campylobacter jejuni and Campylobacter coli was investigated in 64 samples of fresh retail chicken purchased from commercial slaughterhouses located in Brazil. Campylobacter spp. were isolated from 40 (62.5%) of 64 analyzed samples. The strains biotyped according to Lior were classified as C. jejuni biotypes I and II, and C. coli biotypes I and II. The efficiency of different procedures for recovering Campylobacter spp. from chicken carcasses was tested. The enrichment procedure was significantly less effective than direct plating (P < 0.05), detecting 19 of 40 (47.5%) as opposed to 38 of 40 (95%) positive samples. Using direct plating the efficiency of Blasers selective supplement was significantly more effective (P < 0.05) than Skirrows selective supplement. To verify which factors could be affecting Campylobacter spp. growth in enrichment broth, the pH was measured after incubation for 48 h at 42°C and lactobacilli, coliforms, and enterococci were enumerated. Most of the Campylobacter -negative samples presented high levels of indicator microorganisms, which may have hindered the recovery of Campylobacter spp. during the enrichment procedure.

Chicken feet bacteriological quality at 4 steps of technological processing

The production of chicken feet is primarily intended for foreign markets, and there is still no specific legislation in Brazil that determines the quality standard of these products. The bacteriological quality of chicken feet was evaluated as a product for human consumption at different steps of the technological processes. Eighty broiler feet from 20 lots at 4 steps of processing were collected for quantitative analysis, total count of aerobic mesophilic bacteria, and determining the most probable number of coliforms and fecal coliforms. Thirty-eight pools of 15 broiler feet each from 19 lots were used for qualitative analysis and the isolation of Salmonella enterica spp. and Escherichia coli O157:H7. Escherichia coli O157:H7 was not found in any of the samples. Salmonella spp. were isolated in 68% (13/19) of the lots. The Salmonella Schwarzengrund serotype was found in 12 of the 13 lots of positive samples and the Salmonella Anatum and Salmonella Corvallis serotypes were identified in the remaining lot. Processing is effective in reducing contamination by mesophilic bacteria, coliforms, and Salmonella spp. in these products. This work constitutes the first study in Brazil on microbiological quality of chicken feet.

Helicobacter species detection and histopathological changes in stray cats from Niterói, Brazil

Tightly coiled bacteria are common inhabitants of the gastric mucosae of cats and there is considerable debate whether feline helicobacters are commensal or pathogenic organisms. To determine the prevalence of Helicobacter species and gastric histopathological findings, gastric mucosa samples of 56 stray cats were collected from cardia, fundus, body, antrum and pylorus at necropsy. The samples were examined by urease test, cytology, histopathology and culture. Helicobacter species were detected in 53 cats (94.6%) by at least one detection method. Urease test results were positive in gastric samples from 50 (89.3%) cats. Cytological and histological evaluations revealed Helicobacter species in 52 (92.8%) and 50 (89.2%) cats, respectively. Helicobacter species organisms were not cultured from any cat. Regardless of the presence of gastric bacteria, most of the cats had mild to moderate gastritis. These data reveal a high prevalence of Helicobacter species in stray cats from Niterói, Brazil.

Investigação de Campylobacter fetus e Tritrichomonas foetus na mucosa prepucial de touros da região do Médio Paraíba, RJ

Thirty nine breeding bulls from dairy farms (n=9) and beef farms (n=30) located in Medio Paraiba region at Rio de Janeiro - Brazil state were investigated for the presence of Campylobacter fetus and Tritrichomonas foetus. For Campylobacter investigation, smegma samples were examined by culture and prepucial washings were examined by direct immunofluorescence technique (DIF). The prepucial washings were also examined for Tritrichomonas foetus presence by direct examination. C. fetus was identified in 14 samples (35.9 %) by DIF technique and C. fetus subspecies venerealis was isolated from four samples (10.3%). T. foetus was not detected in bull samples. The high frequency of C. fetus observed in bull samples suggests the occurrence of campylobacteriosis among herds which have reproductive problems at the Medio Paraiba region.

Draft Genome Sequences of Salmonella enterica subsp. enterica Serovar Typhimurium Strains Isolated from Chicken and Swine Carcasses in Two Distinct Geographical Regions from Rio de Janeiro State, Brazil

ABSTRACT Salmonella enterica subsp. enterica serovar Typhimurium is a surveyed worldwide serotype with well-characterized genomes for several different strains. In Brazil, very few studies have submitted whole-genome sequences to GenBank. This genome may be useful to analyze the genetic mechanisms comparable to those of other related studies conducted in Brazil and globally.

Detecção de resistência às fluoroquinolonas em Campylobacter isolados de frangos de criação orgânica

Studies have shown that resistance to quinolones in Campylobacter strains is related with Threonine-86-Isoleucine mutation. In order to investigate the presence of this mutation in sensitive and resistant Campylobacter strains to ciprofloxacin and enrofloxacin, the cecal contents of 80 broilers from organic raising chickens, slaughtered under State Inspection Service (S.I.S) of the State of Rio de Janeiro, were collected and tested for the presence of Campylobacter. The determination of ciprofloxacin and enrofloxacin susceptibility was done by disk diffusion and agar dilution methods for determining the Minimum Inhibitory Concentration (MIC). The detection of mutation in Quinolone Resistance Determinant Region (QRDR) in gyrA gene was done by sequencing. Campylobacter was isolated from 100% of the samples, being 68.75% C. jejuni and 31.25% C. coli. By the disk diffusion method, resistance to ciprofloxacin was observed in all isolates and 56.25% of the strains were resistant to enrofloxacin. By agar dilution method, all strains were resistant to ciprofloxacin (MIC ≥ 16μg/mL to ≥ 64μg/mL) and full and intermediate resistance to enrofloxacin was detected in 42.50% (MIC ≥ 4-32μg/mL) and 38.75% (MIC =2μg/mL) of the strains, respectively. Mutation Thr-86-Ile was observed in 100% of the isolates investigated. In addition to this mutation, others no silent mutations (Val-73-Glu, Ser-114-Leu, Val-88-Asp, Ala-75-Asp, Gly-119-Ser, Arg-79-Lys) and silent mutations (His-81-His, Ser-119-Ser, Ala-120-Ala, Phe-99-Phe, Ala-122-Ala, Gly-74-Gly, Ile-77-Ile, Ala-91-Ala, Leu-92-Leu, Val-93-Val, Ile-106-Ile, Thr-107-Thr, Gly-113-Gly, Ile-115-Ile, Gly-110-Gly) were detected. All the enrofloxacin-sensitive strains by the phenotypic methods had the Thr-86 to Ile substitution, which suggests other mechanisms contributing to enrofloxacin resistance in Campylobacter.

Pré-resfriamento na redução de coliformes em carcaças de frango de corte

The objective of this study was to assess the influence of immersion chilling on broiler carcasses contamination by fecal coliforms counting in a poultry slaughterhouse under Federal Sanitary Inspection by individual and three class plan interpretation of the results. Two hundred and forty broiler carcasses, being collected 120 before and 120 after passage thought chillers, were analyzed for coliform counting by plate count technique. The averages of the counts obtained from all carcasses collected before and after immersion chilling were different, with an average reduction of 0.99log10 CFU g-1 of fecal coliforms. Regarding individual results (two class plan) before chilling, 16.7% (20/120) of the carcasses were classified as unacceptable, against 37.5% (45/120) regarding lots results (three class plan). There was a strong association between the acceptability of lots and immersion chilling with an Odds Ratio value of 35.48. It was demonstrated the importance of the official sampling plan and immersion chilling in the production process, being decisive for the acceptance of the chicken carcass lots by the national legislation standards.

Microbiological Findings in Tracheal Wash From Mule Foals With and Without Clinical Evidence of Respiratory Disease

Abstract Respiratory diseases are common in horses; however, there is a lack of information in the literature on respiratory disease affecting mule foals. The objective of this study was to investigate the presence of aerobic bacteria in tracheal wash samples from 20 mule foals up to 6 months of age, with and without clinical evidence of respiratory disease. Samples were collected via endoscopy in two separate occasions and sent for cytology, microbial culture, and PCR for detection of Rhodococcus equi. Based on clinical evidence, 32.5% (13/40) of the samples were obtained from mule foals displaying signs of a respiratory condition, whereas 50% (20/40) of the samples showed cytologic evidence of respiratory tract infection. One hundred percent of samples provided positive cultures with Escherichia coli (45%), Enterococcus (37.5%), and coagulase‐negative Staphylococcus (30%) being the most common bacterial genera isolated. R. equi was not identified in any of the samples. The correlation between isolated bacterial agents and the presence of respiratory infection was not statistically significant. The microorganisms found in the samples may be naturally present in the soil, feces, and environment in which the animals live, presenting a risk of opportunistic respiratory infection. HighlightsAerobic bacteria was investigated in endoscopically obtained tracheal wash of mule foals up to 6 months of age with and without clinical evidence of respiratory disease.Bacterial growth of at least one type of bacteria was obtained in 100% of endoscopically obtained traqueal wash samples from mule foals, and at least two bacteria genera were identified in 87% of the samples.Escherichia coli, Enterococcus, and coagulase‐negative Staphylococcus were the most commonly isolated bacteria found in 45%, 37.5%, and 30.0% of samples, respectively.When frequency of bacteria genera in mule foals with or without cytologic evidence of infection was submitted to statistical analysis, no significant association was detected.

Analysis of findings throughout the cycles of accreditation of veterinary diagnostic testing laboratories for diseases transmitted by products of animal origin

This work describes the frequency of findings registered in the assessment reports of 23 Brazilian animal health laboratories from 2008 to 2016. The study was conducted in consideration of the ISO/IEC 17025 requirements and normative documents issued by the Brazilian Accreditation Body [General Coordination for Accreditation (Cgcre)]. The general performance of these laboratories throughout the evaluation cycles was investigated. The types and number of findings reported in the assessments of these laboratories were analyzed according to the management and technical requirements of ISO/IEC 17025 and Cgcre’s normative documents. The laboratories’ performance was compared in consideration of the number of findings reported in the first and last assessments of each laboratory investigated.

Detecção de resíduos de enrofloxacina por ensaio imunoenzimático e cromatografia líquida acoplada à espectrometria de massas em ovos comerciais de galinha após tratamento

Enrofloxacin is one of the most used antibiotics in the poultry industry and the deposition of residues in poultry products, such as eggs, are of great concern to public health. In Brazilian law there is no standard withdrawal period for enrofloxacin in eggs and there is no Maximum Residue Limit (MRL) established for this antimicrobial in eggs. In this study, (Bioo Scientific(r)) commercial ELISA kit and LC-MS/MS were used to investigate enrofloxacin in eggs of 30 hens pretreated via drinking water at 10mg/kg of enrofloxacin for five days. Six eggs were collected daily and analyzed during treatment and after the end of treatment, for 15 days. Residues obtained maximum levels on the fifth day of treatment, declined gradually and were no longer detected from the ninth day to the end of treatment. Based on the MRL of 100mg/kg established for edible tissues of poultry by Brazillian law and for muscle, fat and skin, by the European Union, after six days of treatment withdrawal, the residue levels were below that limit, with the average of 37.43mg/kg in LC-MS/MS and 14.731mg/kg in ELISA. Within the conditions of this study, a withdrawal period of six days would be more appropriate to use the eggs for human consumption. The values obtained by ELISA for residues in eggs were lower than those obtained in LC-MS/MS for the same sample, however both methods showed statistical agreement. LC-MS/MS is the recommended method by Brazilian legislation for analysis of residues in food, however, according to the results the ELISA kit used can also be applied to the detection of enrofloxacin residues in eggs, with the advantages of speed and simplicity.