High frequency of non-susceptibility to ciprofloxacin in nontyphoid Salmonella recovered from Brazilian broiler chicken.

1. This study evaluated the minimal inhibitory concentration (MIC) of ciprofloxacin and the presence of plasmid-mediated quinolone resistance (PMQR) mechanisms in 97 nontyphoidal Salmonella spp. isolated from broilers and carcases from three different regions in Brazil. The presence of mutations in quinolone resistance determination regions (QRDRS) was investigated in the ciprofloxacin-resistant strain by DNA sequencing.2. Most of the Salmonella spp. (85.6%) had intermediate resistance to ciprofloxacin and only one isolate was resistant. MIC breakpoints ranged from ≤0.03 to 1 µg/ml and 67.0% of the strains had a MIC of 0.25 µg/ml (n=65). Thirteen strains (13.4%) were susceptible to ciprofloxacin with MIC ≤0.06 µg/ml. The qnrB gene was detected in eight isolates with intermediate resistance and in two susceptible strains. The other PMQR genes, qnrA, qnrC, qnrD, qnrS, qnrVC, aac(6')-Ib, qepA, oqxAB and mutations in QRDR were not detected in any strain.3. There was a high frequency of ciprofloxacin intermediate resistant Salmonella from broiler and broiler carcases from Brazil. The presence of these strains in poultry and derived products poses a risk to public health.

Comparison of antimicrobial resistance in thermophilic Campylobacter strains isolated from conventional production and backyard poultry flocks.

1. The aim of this study was to compare the resistance pattern of thermophilic Campylobacter spp. isolated from conventional production (n = 34) and backyard poultry flocks (n = 36) from Rio de Janeiro State, Brazil. The disc diffusion method and statistical tests were used for investigation and analysis of the resistance pattern of Campylobacter spp. isolated from different rearing systems.2. Antimicrobial resistance percentages to amoxycillin with clavulanic acid (AMC), ampicillin (AMP), ceftiofur (CTF), ciprofloxacin (CIP), enrofloxacin (ENO), erythromycin (ERI), gentamicin (GEN) and tetracycline (TET) were 32.4%, 44.1%, 67.6%, 97.1%, 82.4%, 26.5%, 5.9% and 38.2% in conventional production flocks respectively, while the backyard flock's resistance levels were 0.0%, 13.9%, 69.4%, 100.0%, 91.7%, 5.6%, 0.0% and 16.7%, respectively.3. Campylobacter spp. from conventional poultry production was more resistant to AMC, AMO, ERI and TET (P > 0.05) when compared to strains from backyard poultry. A higher frequency of resistance to fluoroquinolones (FLQ), CIP and ENO, was observed in strains from both systems, demonstrating the spread of resistant strains among poultry production environments.

Prevalência e estudo genético de Mycoplasma gallisepticum e M. synoviae em poedeiras comerciais, na região centro-oeste do estado de São Paulo, Brasil / [Prevalence and genetic study of Mycoplasma gallisepticum and M. synoviae strains from commercial laying hens in the Midwest region of Sao Paulo state, Brazil]

O objetivo deste trabalho foi estudar a prevalência de MG e MS e a filogenia das cepas circulantes, comparando-as com outras já descritas em poedeiras comerciais no Brasil. Foram coletados 140 suabes traqueais de poedeiras comerciais com sinais respiratórios em seis granjas da região centro-oeste de São Paulo. As amostras foram avaliadas por PCR, com posterior sequenciamento e análise filogenética das cepas identificadas. Das 140 amostras, 16,4% foram positivas para MG e 68,6% para MS. Houve diferença significativa nas frequências de MG e MS por granja, segundo o teste G de independência (P<0,05). Todas as cepas identificadas de MG e MS de granjas distintas apresentaram similaridade tanto pela lipoproteína para MG quanto pela região 16s rRNA para MS. Neste estudo, foi possível observar altas prevalências dos agentes estudados, sendo a de MS maior que a de MG. Foi detectada infecção mista por MG e MS em 11,4% das amostras e sabe-se que esses micoplasmas podem agir de forma sinérgica, agravando o quadro respiratório. As cepas circulantes identificadas, pela análise das regiões gênicas da lipoproteína para MG e 16S rRNA para MS, são similares em todas as granjas estudadas.(AU)

The aim of this study was to evaluate the prevalence of MG and MS and the phylogeny of the circulating strains, comparing them with others already described in commercial laying hens from Brazil. A total of 140 tracheal swabs were collected from commercial laying hens with respiratory signs in six farms from the western region of São Paulo state. The samples were analyzed by PCR with subsequent sequencing and phylogenetic analysis of the identified strains. From the 140 samples, 68.6% were positive for MS and 16.4% for MG. There was a significant difference in the frequencies of MG and MS per farm according to G Test of independence (P<0.05). All strains identified as MG and MS from distinct farms presented similarity both by lipoprotein to MG and by 16s rRNA region to MS. In this study, it was possible to observe a high prevalence of MS compared to MG. Mixed MG and MS infection was detected in 11.4% of the samples. These mycoplasmas may act synergistically, worsening the respiratory signs. The circulating strains identified by analysis of the lipoprotein for MG and 16S rRNA for MS are similar on all poultry farms studied.(AU)

Caracterização macroscópica e microscópica da pododermatite em relação à classificação industrial de pés de frango para consumo humano / Pododermatitis macroscopic and microscopic characterization related to industrial classification of chicken feet for human consumption

O presente trabalho teve como objetivo identificar e classificar a pododermatite em frangos de corte, comparando as lesões microscópicas com os aspectos macroscópicos utilizados pela inspeção sanitária. Foi realizada coleta de pés de frangos de corte, em matadouro de aves sob inspeção sanitária, após classificação utilizada nos padrões para exportação. Foram coletados 30 pés tipo A, 30 pés tipo B e 33 pés tipo C, escolhidos aleatoriamente dentro de cada grupo. Para análise histopatológica, foram desenvolvidos escores de acordo com a gravidade das lesões, variando de 0 a 2. Foi observada associação (qui-quadrado, P<0,05) entre a classificação macroscópica (A, B e C) e as alterações histológicas (0, 1 e 2). A classificação A diferiu significativamente (ANOVA, Tukey-Kramer, P<0,05) das classificações B e C quanto aos escores histopatológicos observados. Não houve diferença no comprometimento dos pés pelas lesões que justificassem a separação entre os pés classificados em B e C, uma vez que ambos apresentaram delimitação linear das lesões, sugerindo superficialidade e restrição ao epitélio queratinizado. Portanto, sugere-se o aproveitamento dos pés para consumo humano após remoção mecânica do "calo de pé", uma vez que este produto não oferece riscos ao consumidor.(AU)

The aim of this study was to identify and classify pododermatitis in broilers, comparing the microscopic lesions with the macroscopic aspects used by the Sanitary Inspection. Broiler chicken feet were collected at a Poultry slaughterhouse under Sanitary Inspection, after classification according to the exportation standards. The chicken feet were randomly selected in each group, 30 feet type A, 30 feet type B and 33 feet type C. For the histopathological analysis, scores were developed according to the severity of the lesions, varying from 0 to 2. There was association (Chi-square, P< 0.05) between the macroscopic classification (A, B and C) and histological changes (0, 1 and 2). The A classification differed significantly (ANOVA, Tukey-Kramer, P< 0.05) from the B and C classifications regarding the histopathological scores observed. There was no difference in feet lesions that justified the separation between the feet classified in B and C, since both presented a linear delimitation of the lesions, suggesting superficiality and restriction to the keratinized epithelium. Therefore, the use of the feet for human consumption after mechanical removal of the footpad lesions is suggested since the product does not pose risks to the consumer.(AU)

Detecção de resíduos de enrofloxacina por ensaio imunoenzimático e cromatografia líquida acoplada à espectrometria de massas em ovos comerciais de galinha após tratamento / Investigation of enrofloxacin residues through enzyme-linked immunosorbent assay and liquid cromatography-tandem mass spectrometry in eggs from laying hens after treatment

A enrofloxacina é um dos antimicrobianos mais utilizados na avicultura industrial, e a deposição de resíduos em produtos avícolas, como os ovos, são de grande importância para a saúde pública. Na legislação brasileira não existe padronização do período de carência para o seu uso na produção avícola e não há Limite Máximo de Resíduo (LMR) fixado para enrofloxacina em ovos. Neste estudo, foi utilizado o kit de ELISA comercial (Bioo Scientific(r)) e a LC-MS/MS na pesquisa de enrofloxacina em ovos de 30 galinhas tratadas previamente via água de bebida, com 10mg/kg de enrofloxacina, durante cinco dias. Seis ovos foram coletados diariamente e analisados durante o tratamento e após a sua suspensão, durante 15 dias. A deposição de resíduos obteve níveis máximos no quinto dia de tratamento das aves, declinando gradativamente até não ser detectada a partir do nono dia de suspensão do tratamento. Considerando como base o LMR de 100µg/kg fixado pelo Brasil para tecidos comestíveis de aves e pela União Europeia para músculo, gordura e pele, após seis dias de suspensão do tratamento, os níveis de resíduos foram inferiores a esse limite, tendo como médias 37,43µg/kg na LC-MS/MS e 14,731µg/kg no ELISA. Dentro das condições deste estudo, um período de carência de seis dias seria mais adequado para utilização dos ovos para consumo humano. Foram detectados valores de resíduos nos ovos menores no ELISA em relação à LC-MS/MS para a mesma amostra, mas os dois métodos apresentaram concordância estatística entre si. A LC-MS/MS é o teste recomendado pela legislação brasileira para a análise de resíduos em alimentos; entretanto, pelos resultados obtidos, o kit de ELISA utilizado também pode ser aplicado na detecção de resíduos de enrofloxacina em ovos, com as vantagens de rapidez e simplicidade...

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...

The effect of eggshell apex abnormalities on table egg quality during storage in 2 seasons of the year.

Mycoplasma synoviae infection of hens has been associated with problems of eggshell quality called eggshell apex abnormalities (EAA). Little is known about the quality of EAA eggs from a commercial point of view, especially during their storage. The study aimed to examine the differences between EAA and normal eggs during storage under controlled conditions in 2 seasons, summer and winter, by comparing internal and external quality parameters. In a conventional egg production farm with white laying hens of varying ages in the city of Bastos, state of São Paulo, Brazil, 232 eggs were used in the summer season and 400 eggs in the winter season. Half of the eggs had EAA, and the other half were considered normal eggs for each season. The eggs were analyzed at 2, 7, 14, 21, and 28 d after being laid and stored from 24.6 to 25.8°C in summer and from 24 to 25°C in winter. There was no difference (P > 0.05) in the average egg weight between EAA and normal eggs at any studied time point, but in both seasons, the weight loss in EAA eggs was higher than in normal eggs. The losses in Haugh unit scores from the first to the last measurements were approximately 40% regardless of egg type or season of production. In comparing eggshell thickness, only the apices of the EAA eggs were thinner (P < 0.0001) than normal eggs in the summer, but in the winter, the EAA egg apices (P < 0.0001) and sides (P = 0.03) were both thinner. The presence of EAA did not affect the eggshell weight (P > 0.05) or eggshell percentage (P > 0.05). The eggshell strength of the EAA eggs was lower (P < 0.0001) than normal eggs in both the summer (16.57%) and winter (19.86%). The presence of EAA did not affect the internal quality of the egg, but was related to a greater loss of external quality and weight during storage.

Principais alterações no leite por agentes causadores de mastite no rebanho caprino dos estados de Minas Gerais e Rio de Janeiro / Major changes in milk by mastitis causative agents in goat herd from the states of Minas Gerais and Rio de Janeiro, Brazil

A mastite subclínica caprina ocasiona prejuízos econômicos em decorrência do descarte, dos gastos com medidas terapêuticas e da redução da quantidade e qualidade do leite e seus derivados.Nesse estudo, 129 amostras de leite de cabra in natura, provenientes de 11 propriedades foram avaliadas pelo teste da caneca telada, California Mastitis Test (CMT), exame bacteriológico, pesquisa de Mycoplasma spp. e pela determinação dos parâmetros físico-químicos. No teste da caneca telada e no CMT, 3,1% e 4,6% das amostras foram positivas, respectivamente. No exame bacteriológico, 57,4% das amostras foram positivas e o patógeno mais frequente foi Staphylococcus coagulase negativa com 56% das cepas resistentes à penicilina e 100% de sensíveis à gentamicina. Mycoplasma spp. não foi identificado nas amostras. O diagnóstico da mastite subclínica pelo CMT e pelo exame bacteriológico diferiu de forma significativa e não houve associação entre o número de UFC/mL obtidas no exame microbiológico e o resultado do CMT (Qui-quadrado p 0,05). Com base nos resultados obtidos, recomenda-se a associação do exame bacteriológico quando na utilização do CMT para diagnóstico da mastite subclínica caprina.

Major changes in milk by mastitis causative agents in goat herd from the states of Minas Gerais and Rio de Janeiro, Brazil. Goat mastitis causes significant economic losses due to the discarding of milk, costs of drugs and veterinary care, reducing the quantity and quality of milk and dairy products. In this study, 129 raw milk samples from 11 goat farms were investigated by the Tamis test, California mastitis test (CMT), bacteriological exam, presence of Mycoplasma spp. and physicochemical parameters. Seven (4.6%) and four samples (3.1%) were positive by CMT and Tamis test respectively. Bacteriological exam was positive from 57.4% of samples and coagulase-negative Staphylococcus was the most frequent bacteria isolated showing 56% of the strains resistant to penicillin and no resistance to gentamicin. Negative results were obtained from traditional culture as well as by PCR for Mycoplasma spp. The diagnosis of mastitis, the bacteriological exam and the CMT results differed significantly and no association was observed (chi squared, p 0.05). The physicochemical parameters differed significantly (ANOVA, Tukey-Kramer, p < 0.05) among the herds. These results indicate the need to associate microbiological exam when the CMT is used for the diagnosis of goat mastitis.

PCR for the detection of the gene fel a of escherichia coli in broiler condemned for airsacculitis by the Brazilian Federal Sanitary Inspection / PCR na detecção de gene Fel A de Escherichia Coli em frangos de corte condenados por aerossaculite pela Inspeção Sanitária Federal

Collibacillosis is considered one of the major diseases of the modern poultry industry, due to the significant losses it causes. Escherichia coli contributes not only to the disease itself, by causing weight loss of the birds, but also to the increase in carcasses condemnation during slaughter and processing. Detection of virulence factors in E. coli strains of the APEC pathotype contributes to the characterization and pathogenicity of this agent. PCR techniques have been very helpful in the search for genes that encode those virulence factors. This study aimed to detect the gene Fel A of E. coli by PCR and relate its positivity to low weight in broiler flocks with airsacculitis as diagnosed by the health inspection service. The study involved 40 flocks of broilers slaughtered in a single poultry slaughterhouse, under Federal Sanitary Inspection, located in the state of Rio Grande do Sul, Brazil. Three broilers were randomly selected to obtain one "pool" of three tracheas for each PCR. DNA was extracted using phenol-chloroform and amplified using a pair of primers specific to gene Fel A of E. coli. Of the 40 flocks analyzed by PCR, 35% (14/40) were positive for the gene Fel A. PCR was an effective technique for the detection of gene Fel A in broiler flocks. There was a relationship between the presence of the gene Fel A, weight loss, and increase of the airsacculitis rate...

A colibacilose é considerada uma das principais doenças da indústria avícola moderna, devido aos grandes prejuízos econômicos causados. A Escherichia coli contribui não só para a doença em si, levando à perda de peso das aves, bem como para o aumento da taxa condenação de carcaças durante o abate e processamento. A detecção de fatores de virulência de cepas de E. coli do patotipo APEC colabora para a caracterização de sua patogenicidade e as técnicas de PCR têm sido muito úteis na pesquisa de genes que os codificam. Este estudo objetivou diagnosticar E. coli pela detecção o gene Fel A por PCR e relacionar a positividade para este agente com o baixo peso em frangos de corte provenientes de lotes condenados por aerossaculite. Foram estudados 40 lotes de frangos de corte abatidos em um matadouro avícola sob Inspeção Sanitária Federal, localizado no Estado do Rio Grande do Sul. Foram colhidos aleatoriamente 3 frangos e obtidos "pools" de três traqueias em cada um deles para PCR. O DNA foi extraído pelo método de fenol-clorofórmio e amplificado com pares de "primers" específicos para gene Fel A de E. coli. Dos 40 lotes analisados pela PCR, 35% (14/40) foram positivos para o gene Fel A. A PCR foi eficaz para a detecção do gene Fel A em lotes de frangos de corte e houve relação entre a presença do gene Fel A, a queda de peso e aumento na taxa de aerossaculite...

Avian infectious bronchitis and deep pectoral myopathy - a case control study.

Infectious bronchitis is caused by a coronavirus, infectious bronchitis virus (IBV). Infectious bronchitis is an acute and highly contagious disease of economic importance due to the reduction in weight gain observed with infected broilers and the drop in egg quality and production associated with infected laying hens. The presence of deep pectoral myopathy has been associated with IBV variants. This lesion is detected at slaughterhouses and is characterized by paleness and atrophy of the deep pectoral muscle, including necrosis of the region, leading to condemnations of the breast muscle, a valuable meat cut in the market. This work aimed to study the relationship between deep pectoral myopathy and IBV by describing tracheal and muscle lesions and comparing the frequency of IBV detection via reverse-transcription (RT) PCR in muscle, tracheal, and cecal tonsil samples from broilers with and without myopathy. A case-control study was conducted in 40 broiler flocks vaccinated with the Massachusetts strain. The case group consisted of 23 flocks that presented myopathic lesions under sanitary inspection and a control group of 17 flocks without myopathic lesions. The tracheal, cecal tonsil, and supracoracoid muscle (with and without lesions) samples from the 40 broiler flocks were screened by RT-PCR to detect IBV. Histopathology of muscle and tracheal tissue was carried out. Upon microscopic examination, the muscle samples from the case group presented extensive necrosis, intense mononuclear inflammatory infiltration, muscle fiber fragmentation, and fibrotic tissue, confirming myopathy, whereas muscles from the control group showed no alterations. The tracheal samples presented a large number of infiltrated mononuclear inflammatory cells that in some areas formed submucosal nodules. A total of 25 flocks tested IBV positive by RT-PCR: 14 from the case group and 11 from the control group. The IBV was detected by RT-PCR directly in muscle samples. Despite that, the relationship between deep pectoral myopathy and IBV was not established. The higher positive IBV RT-PCR percentage noted in the cecal tonsil samples demonstrates how important the choice of organs is for diagnostic purposes.

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.