Stream and well water samples from two rural communities in Ekiti State, Nigeria: assessment of physicochemical parameters, bacteriological quality and public health significance

Infectious diseases caused by microorganisms are widespread health risks associated with drinking water. This study evaluated the physicochemical parameters and bacteriological quality of the stream and well water using standard protocols. The bacteria were identified by conventional and molecular methods. Antibiotic susceptibility and location of antibiotic resistance markers (ARMs) were determined using disc diffusion and acridine orange, respectively. The highest mean Total Heterotrophic Bacterial Counts (THBC), Total Coliform Counts (TCC) and Faecal Coliform Counts (FCC) from the stream water was 4.3 ± 0.3×106, 8.9 ± 0.0×105, and 3.5 ± 0.1×104 (CFU mL-1), respectively. The well water had mean TCC ranging between 2.8 ± 0.0×103 and 2.1 ± 0.1×104 (CFU mL-1). Six bacterial genera: Staphyloccocus, Pseudomonas, Escherichia, Enterobacter, Klebsiella, and Shigella were isolated. The mean temperature of the water ranged from 26.0 ± 0.3oC to 27.0 ± 0.1oC. The highest mean dissolved oxygen, total hardness, sulphate and magnesium was 24.0 ± 1.0, 40.1 ± 0.8, 11.0± 1.0, and 67.0 ± 1.5 (mg L-1), respectively. The results showed that ≥ 66.7 S. aureus were Levofloxacin and Streptomycin sensitive; between 45.5 and 68.1% of the isolates were Gentamycin and Chloramphenicol resistant, while 81.8% exhibited multidrug resistance. Escherichia coli EcSW3, E. aerogenes EeWW2, K. pneumoniae KpSW3, and S. aureus SaSW had their entire ARMs located on the plasmids with the molecular sizes ˂ 2.027 Kbp. This study showed that the stream and well water harboured bacteria with some ARMs on plasmids, indicating the possibility of horizontal transfer of antibiotic-resistant genes among the bacteria. In addition, it showed the necessity to enlighten the rural populace on the importance of cleaning the surroundings near water sources so as to prevent water-borne diseases.(AU)

Deeper exploration of inflammatory cell populations in milk to monitor udder health in dairy cows

The present study explored the predictive values of milk leukocyte differentials (MLD) as a basis for improving the diagnosis of intramammary infections (IMIs) and subclinical mastitis. Quarter milk samples were collected for bacteriological analysis, quarter somatic cell count (qSCC), and MLD. The MLD were assessed using the cytospin technique, direct microscopic smears, and flow cytometry. The predictive values of each single leukocyte population and useful potential indices that could better reflect immune complexity were also calculated. Changes in the percentage of any leukocyte alone failed to substantially improve the predictive value of qSCC in diagnosing IMIs. Although certain parameters increased the area under the receiver operating characteristic curve (ROC curve) as a result of increased specificity values, a slight reduction in sensitivity was observed. The so-called CD8 complex was a unique parameter which improved both the sensitivity (78.79 %) and the specificity (80.77 %) in IMI diagnosis, resulting in the highest area under the ROC curve (0.87). To diagnose subclinical mastitis, the percentage of macrophages and the sum of the percentage PMNLs and T CD8+ cells divided by the percentage of macrophages showed the highest predictive values (sensitivity = 79.63, specificity = 73.68, and area under the ROC curve = 0.83) in the differentiation of the inflammatory condition status of cows. In conclusion, this study provides further insights into using T CD8+ lymphocytes in diagnosing bovine IMIs, combined with PMNLs and macrophages. The antidromic trend of macrophages vs. PMNLs and T CD8+ lymphocytes due to the increasing qSCCs was crucial to differentiating quarters under both inflammatory and non-inflammatory conditions.

Investigation of Trichomonas gallinae in passerines / Investigação de Trichomonas gallinae em passeriformes

Trichomoniasis, caused by the protozoan Trichomonas gallinae, has as main hosts birds of the Columbidae family, which have a high prevalence of the protozoan without manifesting the disease. The continuous growth of the pigeon population and its cosmopolitan nature mean that today there is a worldwide distribution of this species, being responsible for the distribution and maintenance of the prevalence of trichomoniasis in almost the entire world. The transmission of the disease may be by direct contact, or indirect, through food or water. This indirect route is the reason why such a wide range of bird orders can be infected, very different from columbids such as falconiformes, strigiformes, passerines, piciformes, psittaciformes, gruiformes, galliformes or anseriformes. Thus, the objective of this study was to evaluate the presence of T. gallinae in passerines received at a Wild Animal Screening Center. In order to carry out this study, 300 birds of the order Passerine corresponding to 23 different species were analyzed, received at the Wild Fauna Rehabilitation Center (NURFS) of the Federal University of Pelotas (UFPel), in different seasons of the year between the months of March to October 2021. Samples of swabs from the oropharynx were collected from all individuals, the material was immediately placed in a falcon tube containing Trypticase-Yeast Extract-Maltose (TYM) culture medium and sent to the Laboratory of Protozoology and Entomology (LAPEn), for incubation in a bacteriological growth greenhouse and subsequent identification of the protozoan on a slide in wet mounting under an optical microscope in a 40X objective. Wet mounting on a slide was performed in triplicate and analyzed in its entirety. Of the 300 birds evaluated in in vitro culture for T. gallinae, 25 had inconclusive results and were submitted to PCR analysis, being negative for T. gallinae. Although no positive Passeriformes was found, the monitoring of the occurrence of this protozoan must continue, as it is known that it may easily cause a possible epidemic, leading to losses for the wild fauna that has endangered birds.

Tricomoníase, causada pelo protozoário Trichomonas gallinae, tem como principais hospedeiros aves da família Columbidae, que apresentam alta prevalência do protozoário sem manifestar a doença. O contínuo crescimento da população de pombos e sua natureza cosmopolita fazem com que hoje exista uma distribuição mundial desta espécie, sendo responsável pela distribuição e manutenção da prevalência da tricomoníase em quase todo o mundo. A transmissão da doença pode ser por contato direto, ou indireto, por meio de alimentos ou água. Essa rota indireta é a razão pela qual uma gama tão ampla de ordens de aves pode ser infectada, muito diferente dos columbídeos, como falconiformes, strigiformes, passeriformes, piciformes, psittaciformes, gruiformes, galliformes ou anseriformes. Desta forma, o objetivo desse estudo foi avaliar a presença de T. gallinae em passeriformes recebidos em um Centro de Triagem de Animais Silvestres. Para a realização deste estudo foram analisados 300 aves por conveniência da ordem Passeriforme correpondente a 23 espécies diferentes recebidos no Núcleo de Reabilitação da Fauna Silvestres (NURFS) da Universidade Federal de Pelotas (UFPel), em estações distintas do ano comprendidas entre os meses de março a outubro de 2021. De todos os indivíduos foram colhidas amostra de suabe da orofaringe, o material foi imediatamente acondicionado em tubo falcon contendo meio de cultura Tripticase-Yeast Extract-Maltose (TYM) e encaminhado ao Laboratório de Protozoologia e Entomologia (LAPEn), para incubação em estufa de crecimento bacteriologico e posterior identificação em lamina em montagem úmida no microscópio óptico em objetivo de 40X do protozoário. A montagem úmida em lamina, foi feita em triplicata e analisada em sua totalidade. Das 300 aves avaliadas no cultivo in vitro para T. gallinae, 25 tiveram resultado inconclusivo e foram sumetidas a análise de PCR, sendo negativas para T. gallinae. Embora não tenha sido encontrado nenhum Passeriformes positivo, o monitoramento da ocorrência desse protozoário deve continuar, pois sabe-se que ele pode causar uma possível epidemia facilmente, levando a perdas para a fauna silvestre que possuí aves ameaçadas de extinção.

Meningoencephalitis of unknown origin in dogs

Background: Meningoencephalitis of unknown origin (MUO) is a critical cause of neurological disorders in dogs, mainly affecting small young individuals. Its symptomatology is varied and depends on the affected neuroanatomic region. The ante mortem diagnosis of this condition is uncertain, being achieved by discarding other conditions and often occurring definitively only by performing a necropsy. Thus, this study aims to report 2 cases of meningoencephalitis, one necrotizing and the other granulomatous in dogs. Cases: Case 1. A 3-year-old, Shih Tzu bitch with a body weight of 4 kg, showing proprioceptive ataxia, behavior of walking in circles, and evolving rapidly to non-ambulatory paresis, was treated. The neurological examination showed a posture of decerebrate stiffness and absence of withdrawal reflex and proprioception, suggesting brainstem injury. Laboratory tests showed mild neutrophilia and lymphopenia, while the rapid test for distemper was non-reactive. The cerebrospinal fluid (CSF) analysis showed lymphocytic pleocytosis, and the PCR tests of the CSF, blood, and urine for the detection of infectious diseases were negative, as well as the culture. With no improvement in clinical condition and exams showing a progressive degenerative condition unresponsive to available treatments, the tutor opted for euthanasia of the patient. The subsequently requested necropsy confirmed the diagnosis of granulomatous meningoencephalitis. Case 2. This case refers to a 1-year-and-5-month-old male Maltese breed weighing 4.8 kg. This animal presented walking in circles behavior and loss of vision for a week, with signs worsening rapidly. In the neurological evaluation, the patient presented sensitivity in the middle ear, difficulty opening the mouth, hearing deficit in the right ear, blindness in the right eye, a proprioceptive deficit in the right anterior limb, and head pressing. Laboratory tests showed nonregenerative anemia and mild lymphopenia. After 1 day of hospitalization, the patient showed worsening clinical condition, with obstruction, absence of facial and auricular sensitivity, and nasal stimulus. In addition, onset of generalized seizures was observed; therefore, CSF was analyzed, which did not present significant alterations except for detecting reactive lymphocytes. The bacteriological culture of CSF resulted in no bacterial growth. In addition, the same neurological PCR panel performed for the previous patient was negative. After 5 days of intensive care, the patient presented a cardiorespiratory arrest and died. The subsequently requested necropsy confirmed the diagnosis of necrotizing meningoencephalitis. Discussion: The 2 reported cases confirm that MUO should be considered during the differential diagnoses of patients with neurological alterations. It is known that small-breed dogs are predisposed to these diseases. Laboratory tests and medical imaging are crucial for clinical guidance, helping to discard other neurological pathologies, especially those due to bacterial, fungal, and/or viral agents. However, definitive diagnosis of MUO can only be performed through necropsy and histopathological analysis. For the reported cases, CSF analysis, neurological PCR panel for detecting possible infectious agents, and bacterial culture were essential to rule out other possible causes of meningoencephalitis. Unfortunately, MUO includes progressive neurological disorders causing the patient's death.

Wooden sticks for plaque streaking and microbiological inoculation might be more cost- effective, but is its large scale use feasible? Quality control methods and proof of concept / Métodos de controle de qualidade e prova de conceito para a utilização de palitos de madeira em microbiologia podem ser mais econômicas, mas seu uso em larga escala é viável? Método de controle de qualidade e prova de conceito

The loop is a material classically used in the laboratory for the purpose of plate streaking and handling biological materials. However, metal loops techniques might be time consuming, considering the amount of time spent to guarantee its cooling process through each inoculation. Furthermore, plastic loops may also represent environmental issues during its production and discard process and can also represent higher costs for the laboratory. Thus, in situations of limited resources, even the simplest materials can be restricted due to logistical and budgetary issues, especially in developing countries. Inspired by demands like these, facing an occasional shortage of supply of laboratory plastic handles, we hereby present a quality control for sterilization methods and cost-effectiveness studies towards the use of wooden sticks in a Latin American country and we discuss the possibility of the large-scale use of this technique.

A alça calibrada é um material usado classicamente em laboratório para fins de inoculação em placas e manuseio de materiais biológicos. No entanto, as técnicas de alças metálicas podem consumir muito tempo, considerando a quantidade de tempo gasto para garantir seu processo de resfriamento a cada inoculação. Além disso, alças de plástico também podem representar questões ambientais durante o processo de produção e descarte e também podem representar custos mais altos para o laboratório. Assim, em situações de recursos limitados, até os materiais mais simples podem ser restringidos devido a questões logísticas e orçamentárias, especialmente nos países em desenvolvimento. Inspirados por demandas como essas, diante de uma escassez ocasional de suprimentos de alças de plástico de laboratório, apresentamos um controle de qualidade para métodos de esterilização e estudos de custo-efetividade para o uso de varas de madeira em um país latino-americano e discutimos a possibilidade de grande uso em escala dessa técnica.

Wooden sticks for plaque streaking and microbiological inoculation might be more cost- effective, but is its large scale use feasible? Quality control methods and proof of concept / Métodos de controle de qualidade e prova de conceito para a utilização de palitos de madeira em microbiologia podem ser mais econômicas, mas seu uso em larga escala é viável? Método de controle de qualidade e prova de conceito

The loop is a material classically used in the laboratory for the purpose of plate streaking and handling biological materials. However, metal loops techniques might be time consuming, considering the amount of time spent to guarantee its cooling process through each inoculation. Furthermore, plastic loops may also represent environmental issues during its production and discard process and can also represent higher costs for the laboratory. Thus, in situations of limited resources, even the simplest materials can be restricted due to logistical and budgetary issues, especially in developing countries. Inspired by demands like these, facing an occasional shortage of supply of laboratory plastic handles, we hereby present a quality control for sterilization methods and cost-effectiveness studies towards the use of wooden sticks in a Latin American country and we discuss the possibility of the large-scale use of this technique.(AU)

A alça calibrada é um material usado classicamente em laboratório para fins de inoculação em placas e manuseio de materiais biológicos. No entanto, as técnicas de alças metálicas podem consumir muito tempo, considerando a quantidade de tempo gasto para garantir seu processo de resfriamento a cada inoculação. Além disso, alças de plástico também podem representar questões ambientais durante o processo de produção e descarte e também podem representar custos mais altos para o laboratório. Assim, em situações de recursos limitados, até os materiais mais simples podem ser restringidos devido a questões logísticas e orçamentárias, especialmente nos países em desenvolvimento. Inspirados por demandas como essas, diante de uma escassez ocasional de suprimentos de alças de plástico de laboratório, apresentamos um controle de qualidade para métodos de esterilização e estudos de custo-efetividade para o uso de varas de madeira em um país latino-americano e discutimos a possibilidade de grande uso em escala dessa técnica.(AU)

Maintenance of quality of eggs submitted to treatment with propolis extract and sanitizers

This study aimed to evaluate the internal and microbiological quality of eggs submitted to different types of shell treatments. One hundred and forty-four fresh red eggs were distributed in a scheme of 4 treatments (no washing; washing and immersion in chlorine; washing and immersion in peracetic acid; and washing and spraying of propolis extract) x 5 storage periods (7, 14, 21, 28, and 35 days), stored at 25°C, in each period 6 eggs per treatment were analyzed. The parameters to assess were: weight loss; shell weight; yolk weight; albumen weight, yolk diameter; yolk height; albumen height and Haugh unit (HU). The microbiological quality of eggs was evaluated at 35 days through analysis for aerobic mesophilic bacteria, thermotolerant coliforms, Staphylococcus spp., Salmonella spp. and molds and yeasts. The treatment with propolis extract was the only effective one to maintain the high HU quality of the eggs until 21 days of storage at 25°C and was effective against microbiological contamination of all bacterial groups. The results presented showed greater effectiveness of the propolis extract for maintenance of internal and microbiological quality of eggs, it can be an alternative product to chemical sanitizers.(AU)

Serum calprotectin levels in dogs with diarrhea

Background: Diarrhea induced by infectious factors may lead to significant health problems in dogs. Canine parvovirus(CPV), canine coronavirus (CCV), canine distemper virus (CDV), Giardia spp., Escherichia coli (E. coli), and Salmonella spp. are the important infectious agents that may induce diarrhea in dogs. The present study aimed to investigatethe effect of CPV, CCV, CDV, Giardia spp., E. coli, and Salmonella spp. infections on the change in serum calprotectin(Calp) concentration.Materials, Methods & Results: A total of 30 dogs were enrolled in the study. The study dogs were divided into 3 groups.Healthy animals as confirmed by clinical examination and animals negative for the specified pathogens were placed inGroup 1. Animals infected by one or more agents, including CPV, CCV, CDV, and Giardia spp., but negative for E. coli orSalmonella spp. were placed in Group 2. Finally, animals positive for E. coli or Salmonella spp. and infected or not infectedby one or more agents, including CPV, CCV, CDV, and Giardia spp., were placed in Group 3. Stool samples and rectaland conjunctival swab samples were collected to investigate the etiologic agents that induced diarrhea. Blood samples werecollected through vena cephalica antebrachii for hematological and biochemical examinations. The samples were obtainedvia routine clinical examinations at the Prof. Dr. Servet Sekin outpatient clinic at Dicle University Veterinary Faculty. CPV,CCV, CDV, and Giardia spp. diagnoses were made based on immunochromatographic test kits. The bacteriological analysisof stool samples was used to diagnose E. coli and Salmonella spp. infection...

Serum calprotectin levels in dogs with diarrhea

Background: Diarrhea induced by infectious factors may lead to significant health problems in dogs. Canine parvovirus(CPV), canine coronavirus (CCV), canine distemper virus (CDV), Giardia spp., Escherichia coli (E. coli), and Salmonella spp. are the important infectious agents that may induce diarrhea in dogs. The present study aimed to investigatethe effect of CPV, CCV, CDV, Giardia spp., E. coli, and Salmonella spp. infections on the change in serum calprotectin(Calp) concentration.Materials, Methods & Results: A total of 30 dogs were enrolled in the study. The study dogs were divided into 3 groups.Healthy animals as confirmed by clinical examination and animals negative for the specified pathogens were placed inGroup 1. Animals infected by one or more agents, including CPV, CCV, CDV, and Giardia spp., but negative for E. coli orSalmonella spp. were placed in Group 2. Finally, animals positive for E. coli or Salmonella spp. and infected or not infectedby one or more agents, including CPV, CCV, CDV, and Giardia spp., were placed in Group 3. Stool samples and rectaland conjunctival swab samples were collected to investigate the etiologic agents that induced diarrhea. Blood samples werecollected through vena cephalica antebrachii for hematological and biochemical examinations. The samples were obtainedvia routine clinical examinations at the Prof. Dr. Servet Sekin outpatient clinic at Dicle University Veterinary Faculty. CPV,CCV, CDV, and Giardia spp. diagnoses were made based on immunochromatographic test kits. The bacteriological analysisof stool samples was used to diagnose E. coli and Salmonella spp. infection...(AU)

Eficácia in sílico do florfenicol no tratamento de pododermatite infecciosa por Fusobacterium necrophorum em ovelhas / In sílico efficacy of florfenicol in the treatment of Fusobacterium necrophorum infectious pododermatitis in sheep

Objetivou-se comparar o efeito in silico do florfenicol nas doses de 20 e 30 mg/Kg em ovinos pelas vias intravenosa (IV) e intramuscular (IM), usando a modelagem PK/PD. Realizou-se uma simulação de Monte Carlo com base nos dados de concentração plasmática de um estudo publicado anteriormente. Calculou-se a área sob a curva (ASC) e as taxas de eficácia do florfenicol para os efeitos bacteriostático, bactericida e de erradicação bacteriológica. A dose de 20 mg/Kg IV demonstrou efeitos de erradicação de 100, 93 e 0% para CIM de 0,5, 1 e acima, respectivamente. O efeito bacteriostático foi de 99 e 90% para CIM de 4 e 2 µg/ml, enquanto o bactericida foi de 14% para CIM de 2 µg/ml. A dose de 30 mg/Kg IV apresentou 100% de erradicação para CIM de 1 µg/mL e 100% de efeito bactericida para CIM de 2 µg/mL. Há 100% de efeito bacteriostático em CIM de 4 µg/ml. As doses de 20 e 30 mg/Kg IM mostraram 100% de erradicação para CIM até 1 µg/mL e 0% para CIM maiores. O efeito bacteriostático foi mantido em 100% para uma CIM de 4 µg/mL em ambas as doses. Este estudo mostra o efeito de erradicação bacteriológica do florfenicol nas doses de 20 e 30 mg/Kg, IV e IM. Recomenda-se que seja feito um estudo de eficácia in vivo com a dose de 30mg/Kg IM em ovinos infectados por F. necrophorum com MIC superior a 2 µg/mL.

We aimed to compare the in silico effect of florfenicol at doses of 20 and 30 mg/Kg in sheep by intravenous (IV) and intramuscular (IM) routes, using PK/PD modeling. We performed a Monte Carlo simulation based on plasma concentration data from a previously published study. We calculated the area under the curve (AUC) and the efficacy rates of florfenicol to bacteriostatic, bactericidal, and bacteriological eradication effects. The dose of 20 mg/Kg IV demonstrated 100, 93, and 0% eradication effects for MICs of 0.5, 1, and above, respectively. The bacteriostatic effect was 99 and 90% for MIC of 4 and 2 µg/ml, while the bactericide was 14% for MIC of 2 µg/ml. The 30 mg/Kg IV dose showed 100% eradication for MIC of 1 µg/mL and 100% bactericidal effect for MIC of 2 µg/mL. There is a 100% of bacteriostatic effect at MIC of 4 µg/ml. Doses of 20 and 30 mg/Kg IM showed 100% eradication for MIC up to 1 µg/mL and 0% for MIC above. The bacteriostatic effect was maintained at 100% for a MIC of 4 µg/mL at both doses. This study shows the bacteriological eradication effect of florfenicol at doses of 20 and 30 mg/Kg, IV, and IM. Therefore, we recommend an in vivo efficacy study with a dose of 30mg/Kg IM in sheep infected with F. necrophorum with MIC greater than two µg/mL.

Thoracic spinal epidural empyema in a cat

A 2-month-old male domestic shorthair cat was referred to a private veterinary clinic with a history of dyspnea and inappetence. At the clinical examination, the cat was in poor body condition, with hyperthermia and abdominal distension. No major abnormalities were observed in the thoracic radiographs or abdominal ultrasound. A complete blood count revealed leukocytosis with neutrophilia. Although no clinical diagnosis was made, the cat received support therapy, but went into cardiac arrest and died. The cat was submitted for necropsy and the main gross finding was two white nodules on the ventral surface of the thoracic vertebrae (from T1 to T4). At the cut surface, the nodules were friable and filled with yellow exudate. The epidural space of the thoracic region was filled with yellowish viscous material. Histologically, the vertebrae were partially replaced by abscess formation characterized by a necrotic center with degenerate neutrophils, surrounded by fibrous connective tissue. The epidural space was filled with degenerate neutrophils, necrotic debris, fibrin, and intralesional colonies of gram-negative short rod-shaped to coccobacillary bacteria. Bacteriologic culture yielded Pasteurella multocida. This paper describes the gross, histological, and bacteriological features of a rare case of spinal epidural empyema caused by Pasteurella multocida in a cat.(AU)

Farm microbiological milk culture: study case on cow performance, financial and economic aspects / Cultivo microbiológico de leite na fazenda: estudo de caso sobre desempenho de vacas, aspectos financeiros e econômicos

This study evaluated the use of microbiological culture of milk from cows with clinical mastitis (CM), and the performance and economic results after implementing this procedure. The 18-month data were obtained from a farm in Minas Gerais State, Brazil, with an average daily production of 23.1 L of milk from cows milked twice daily under a semi-intensive regime. After a case of CM was identified, a milk sample from the affected quarter was collected and sent to the farm's laboratory. First, a bi-plate containing selective growth medium was used for isolation of Gram-positive and negative bacteria (Plate 1). Subsequently, a tri-plate with selective growth medium was used for isolation of Gram-positive and negative bacteria, and bacteria of the genus Streptococcus (Plate 2). Finally, a tri-plate containing three chromogenic culture media capable of identifying 18 bacterial species was used (Plate 3). Clinical cases of mastitis were treated once a day based on the results of the microbiological culture. Two economic scenarios were evaluated (scenarios 1 and 2). Scenario 1 compared the situation if all cases of CM were treated (not using on-farm culture) vs. the use of on-farm culture (real data) and the generated savings for one year. Data from 1,582 lactations of 1,227 cows were evaluated, with 1,917 cases of CM from 636 cows recorded. The average annual incidence of CM was 48.2%. Of all cases evaluated, 76.8% were classified as grade 1 mastitis; 20% as grade 2, and 3.2% as grade 3. The incidence of new clinical cases of mastitis was 4.17% per month. From the samples analyzed on the three plates, 27.8% of the cases received a recommendation to not be treated and 72.2% received a recommendation of treatment. However, only 18.6% were not treated, making a total of 81.4% treated cases. Of the clinical cases that did not receive intramammary antibiotic therapy, 84.3% had a clinical cure. Conversely, the clinical cure rate reached 84% for the cases that received intramammary antibiotic therapy. In scenario 1, the total operational cost of the clinical case decreased by 10.3% after the implementation of the on-farm culture, with an 18.4% reduction in the use of antibiotics. In scenario 2, there was a 5.5% reduction in the cost of the clinical case and an 11.8% reduction in the use of antibiotics. Thus, the implementation of on-farm culture and the applied methodology, enhanced treatment accuracy of CM cases, reducing the total operating cost of the case and the use of antibiotics on the farm.

O objetivo deste estudo foi avaliar a utilização da cultura microbiológica de leite de vacas com mastite clínica (MC), os resultados zootécnicos e econômicos após implementação desse procedimento. Dados de 18 meses foram obtidos em uma fazenda em Minas Gerais com produção média de 23,1 L de leite/vaca/dia, em duas ordenhas diárias em regime semi-intensivo. Após a identificação da MC, uma amostra de leite do quarto afetado foi coletada e encaminhada para laboratório da fazenda. Primeiramente foi utilizada placa bipartida em meio de cultura seletivo para crescimento de bactérias Gram-positivas e negativas (Placa 1), seguindo-se para a utilização da placa tri-partida com meios de cultura seletivos para crescimento de bactérias Gram-positivas e negativas e bactérias do gênero Streptococcus (Placa 2) e posteriormente para placa tripartida, contendo três meios de cultura cromogênicos, capazes de identificar 18 espécies bacterianas (Placa 3). Foram avaliados dados de 1.227 vacas em 1.582 lactações, sendo registrados 1.917 casos de MC. Os casos clínicos de mastite foram tratados uma vez ao dia com base nos resultados da cultura microbiológica. Foram realizadas duas avaliações financeiras (cenário 1 e 2). O cenário 1 comparou a situação se todos os casos de MC fossem tratados (ausência de cultura na fazenda), comparado com a utilização da cultura na fazenda (dados reais) e as economias de recursos geradas durante o período de um ano. O cenário 2 utilizou os resultados reais após a economia de recursos gerados pela cultura, seguindo a recomendação ideal de tratamento. Foram avaliados 1.917 casos de MC (636 animais), totalizando incidência média anual de 48,2%. Do total dos casos avaliados, 76,8% foram classificados como mastite grau 1; 20% grau 2 e 3,2% grau 3. A incidência de novos casos clínicos de mastite foi de 4,17% ao mês. Das amostras analisadas nas três placas, 27,8% dos casos receberam a recomendação de não serem tratados e 72,2% de tratamento. Entretanto, apenas 18,6% realmente não foram tratados, totalizando 81,4% de tratamentos. Dos casos clínicos que não receberam antibioticoterapia intramamária, 84,3% apresentaram cura clínica. Já os casos que passaram por antibioticoterapia intramamária, a taxa de cura clínica foi de 84,0%. No primeiro cenário, após a implementação da cultura na fazenda, o custo operacional total do caso clínico reduziu em 10,3%, com redução de 18,4% de utilização de antibióticos. Já no segundo cenário, houve redução de 5,5% no custo do caso clínico e redução de 11,8% na utilização de antibióticos. A implementação da cultura na fazenda e a metodologia aplicada, promoveu maior assertividade dos tratamentos dos casos de MC, com redução do custo operacional total do caso e da utilização de antibióticos na propriedade.

Isolation and molecular identification of Mycobacterium bovis in tissue lesions of cattle slaughtered in slaughterhouses located in the State of Ceará / [Isolamento e identificação molecular de Mycobacterium bovis em lesões teciduais de bovinos abatidos em matadouros-frigoríficos localizados no estado do Ceará]

The objective of this research was to identify Mycobacterium bovis in lesions suggestive of tuberculosis in bovine carcasses in the State of Ceará, by means of bacteriological and molecular diagnostic tests. Between August 2017 and January 2019, the State inspection service (SIE) inspected 59,512 cattle, of which 7.4% (44 / 59,512) presented suggestive lesions. Of these animals, 68 samples were sent, of which 4.5% (31/68) located in the lung, 2.9% (20/68) in lymph nodes, 2.0% (14/68) in the liver, and 0.4% in the carcass (3/68). When performing bacteriological isolation, 15.9% (7/44) of bovines showed colony growth in the samples. The smears of the isolates were submitted to Zielh-Neelsen staining and all confirmed acid-fast bacilli. The polymerase chain reaction identified all isolates 100% (7/7) as M. bovis. The association of diagnostic techniques allowed to identify the presence of the agent in the State and the molecular analysis proved to be a beneficial technique in the monitoring of bovine tuberculosis and can be used as an auxiliary method in the bovine tuberculosis control and eradication program in the State of Ceará.

O objetivo do trabalho foi pesquisar Mycobacterium bovis em lesões sugestivas de tuberculose nas carcaças de bovinos no estado do Ceará, por meio dos testes de diagnóstico bacteriológico e molecular. Entre agosto de 2017 e janeiro de 2019, o Serviço de Inspeção Estadual (SIE) inspecionou 59.512 bovinos; destes, 7,4% (44/59.512) apesentaram lesões sugestivas. Desses animais foram enviadas 68 amostras, das quais 4,5% (31/68) estavam localizadas no pulmão, 2,9% (20/68) nos linfonodos, 2,0% (14/68) no fígado e 0,4% (3/68) na carcaça. Ao realizar o isolamento bacteriológico, 15,9% (7/44) dos bovinos evidenciaram crescimento de colônias nas amostras. Os esfregaços dos isolados foram submetidos à coloração de Zielh-Neelsen e todos eles confirmaram bacilo álcool-ácido resistente. A reação em cadeia da polimerase identificou todos os isolados, 100% (7/7), como M. bovis. A associação das técnicas de diagnóstico permitiu identificar a presença do agente no estado, e a análise molecular demonstrou ser uma técnica benéfica no monitoramento da tuberculose bovina, podendo ser utilizada como um método auxiliar no programa de controle e erradicação da tuberculose bovina no estado do Ceará.

Species identification and antimicrobial susceptibility profile of bacteria associated with cow mastitis in southern Brazil / Identificação de espécies e perfil de suscetibilidade aos antimicrobianos de bactérias associadas à mastite bovina no sul do Brasil

Bovine mastitis is the most common disease in dairy cattle and responsible for economic losses in the milk industry. The present study aimed to identify the main species and to evaluate the antimicrobial susceptibility of bacterial isolates from cow herds with mastitis in dairy farms from southern Brazil. A total of 107 milk samples were collected from different cow herds in one important dairy producing region in southern Brazil, including farms located in ten cities from the Northeast region in the Rio Grande do Sul state. Bacterial strains were isolated and submitted to presumptive identification by classical bacteriological methods. Bacterial species were also identified by MALDI-TOF MS and antimicrobial susceptibility testing was performed with 12 antimicrobials commonly used in dairy farms. Fifty-one bacterial strains were isolated and the presumptive identification demonstrated the occurrence of Staphylococcus spp. (82.3%), Bacillus spp. (3.9%), Klebsiella spp. (3.9%), Streptococcus spp. (3.9%), Corynebacterium sp. (2%), Enterococcus sp. (2%) and Serratia sp. (2%). Forty-one isolates were successfully identified in the MALDI-TOF analysis, including 35 isolates from eleven different bacterial species. Importantly, there were eight different Staphylococcus species, with a high frequency of Staphylococcus chromogenes (48.6%) and Staphylococcus aureus (20%). Overall, bacterial isolates demonstrated resistance to penicillin (46.3%), tetracycline (39%), amoxicillin (36.6%), ampicillin (34.1%) and sulfamethoxazole/trimethoprim (31.7%). Enrofloxacin was the unique antimicrobial that all isolates were susceptible. In addition, there were six multidrug resistant isolates (five S. chromogenes and one S. aureus). This study highlights that bacterial pathogens with resistance to several antimicrobials were identified in cows from dairy farms in a very important milk producing region located in southern Brazil. Microbial identification of the bovine mastitis pathogens and determination of the antimicrobial profile is necessary for the rational use of the medicines.

A mastite bovina é a doença mais comum em gado leiteiro e responsável por perdas econômicas na indústria de laticínios. O presente estudo teve como objetivo identificar as principais espécies e avaliar a suscetibilidade antimicrobiana de isolados bacterianos de rebanhos bovinos com mastite em fazendas leiteiras no sul do Brasil. Um total de 107 amostras de leite foram coletadas em diferentes rebanhos bovinos em uma importante região produtora de leite do sul do Brasil, incluindo fazendas localizadas em 10 cidades da região Nordeste do estado do Rio Grande do Sul. As cepas bacterianas foram isoladas e submetidas à identificação presuntiva por métodos bacteriológicos clássicos. A identificação bacteriana foi confirmada por MALDI-TOF MS e o teste de sensibilidade antimicrobiana foi realizado com antimicrobianos comumente usados em fazendas leiteiras. Cinquenta e uma cepas bacterianas foram isoladas e a identificação presuntiva demonstrou a ocorrência de Staphylococcus spp. (82,3%), Bacillus spp. (3,9%), Klebsiella spp. (3,9%), Streptococcus spp. (3,9%), Corynebacterium sp. (2%), Enterococcus sp. (2%) e Serratia sp. (2%). Os 41 isolados foram identificados com sucesso na análise MALDI-TOF, incluindo 35 isolados de onze espécies bacterianas diferentes. É importante ressaltar que houve a ocorrência de oito espécies diferentes de Staphylococcus, com alta frequência de Staphylococcus chromogenes (48,6%) e Staphylococcus aureus (20%). No geral, os isolados bacterianos tiveram alta resistência à penicilina (46,3%), tetraciclina (39%), amoxicilina (36,6%), ampicilina (34,1%) e sulfametoxazol/trimetoprima (31,7%). A enrofloxacina foi o único antimicrobiano que todos os isolados foram suscetíveis. Além disso, havia seis isolados multirresistentes (cinco S. chromogenes e um S. aureus). Este estudo destaca que os patógenos bacterianos com resistência aos antimicrobianos estão presentes em fazendas leiteiras de subsistência em uma importante região produtora no sul do Brasil. É necessário o monitoramento constante dos patógenos da mastite bovina e a determinação de seu perfil antimicrobiano para o uso racional dos medicamentos.

Species identification and antimicrobial susceptibility profile of bacteria associated with cow mastitis in southern Brazil / Identificação de espécies e perfil de suscetibilidade aos antimicrobianos de bactérias associadas à mastite bovina no sul do Brasil

Bovine mastitis is the most common disease in dairy cattle and responsible for economic losses in the milk industry. The present study aimed to identify the main species and to evaluate the antimicrobial susceptibility of bacterial isolates from cow herds with mastitis in dairy farms from southern Brazil. A total of 107 milk samples were collected from different cow herds in one important dairy producing region in southern Brazil, including farms located in ten cities from the Northeast region in the Rio Grande do Sul state. Bacterial strains were isolated and submitted to presumptive identification by classical bacteriological methods. Bacterial species were also identified by MALDI-TOF MS and antimicrobial susceptibility testing was performed with 12 antimicrobials commonly used in dairy farms. Fifty-one bacterial strains were isolated and the presumptive identification demonstrated the occurrence of Staphylococcus spp. (82.3%), Bacillus spp. (3.9%), Klebsiella spp. (3.9%), Streptococcus spp. (3.9%), Corynebacterium sp. (2%), Enterococcus sp. (2%) and Serratia sp. (2%). Forty-one isolates were successfully identified in the MALDI-TOF analysis, including 35 isolates from eleven different bacterial species. Importantly, there were eight different Staphylococcus species, with a high frequency of Staphylococcus chromogenes (48.6%) and Staphylococcus aureus (20%). Overall, bacterial isolates demonstrated resistance to penicillin (46.3%), tetracycline (39%), amoxicillin (36.6%), ampicillin (34.1%) and sulfamethoxazole/trimethoprim (31.7%). Enrofloxacin was the unique antimicrobial that all isolates were susceptible. In addition, there were six multidrug resistant isolates (five S. chromogenes and one S. aureus). This study highlights that bacterial pathogens with resistance to several antimicrobials were identified in cows from dairy farms in a very important milk producing region located in southern Brazil. Microbial identification of the bovine mastitis pathogens and determination of the antimicrobial profile is necessary for the rational use of the medicines.(AU)

A mastite bovina é a doença mais comum em gado leiteiro e responsável por perdas econômicas na indústria de laticínios. O presente estudo teve como objetivo identificar as principais espécies e avaliar a suscetibilidade antimicrobiana de isolados bacterianos de rebanhos bovinos com mastite em fazendas leiteiras no sul do Brasil. Um total de 107 amostras de leite foram coletadas em diferentes rebanhos bovinos em uma importante região produtora de leite do sul do Brasil, incluindo fazendas localizadas em 10 cidades da região Nordeste do estado do Rio Grande do Sul. As cepas bacterianas foram isoladas e submetidas à identificação presuntiva por métodos bacteriológicos clássicos. A identificação bacteriana foi confirmada por MALDI-TOF MS e o teste de sensibilidade antimicrobiana foi realizado com antimicrobianos comumente usados em fazendas leiteiras. Cinquenta e uma cepas bacterianas foram isoladas e a identificação presuntiva demonstrou a ocorrência de Staphylococcus spp. (82,3%), Bacillus spp. (3,9%), Klebsiella spp. (3,9%), Streptococcus spp. (3,9%), Corynebacterium sp. (2%), Enterococcus sp. (2%) e Serratia sp. (2%). Os 41 isolados foram identificados com sucesso na análise MALDI-TOF, incluindo 35 isolados de onze espécies bacterianas diferentes. É importante ressaltar que houve a ocorrência de oito espécies diferentes de Staphylococcus, com alta frequência de Staphylococcus chromogenes (48,6%) e Staphylococcus aureus (20%). No geral, os isolados bacterianos tiveram alta resistência à penicilina (46,3%), tetraciclina (39%), amoxicilina (36,6%), ampicilina (34,1%) e sulfametoxazol/trimetoprima (31,7%). A enrofloxacina foi o único antimicrobiano que todos os isolados foram suscetíveis. Além disso, havia seis isolados multirresistentes (cinco S. chromogenes e um S. aureus). Este estudo destaca que os patógenos bacterianos com resistência aos antimicrobianos estão presentes em fazendas leiteiras de subsistência em uma importante região produtora no sul do Brasil. É necessário o monitoramento constante dos patógenos da mastite bovina e a determinação de seu perfil antimicrobiano para o uso racional dos medicamentos.(AU)

Bacteria isolated from free-range chicken (Gallus gallus domesticus) eggs sold in semiarid conditions and its one health impact

Background: Food contamination is an important and growing public health concern due to the risk of foodborne illnesses.In this context, the egg, consumed all over the world, stands out. This food has been pointed out as a carrier of severalbacteria, causing outbreaks of food toxinfection. The production of free-range chicken (Gallus gallus domesticus) eggs hasbeen an alternative for generating income for producers in Brazil; however, there is no monitoring of the sanitary qualityof this product. The objective of this study was to identify microorganisms in 128 free-range chicken eggs sold in openmarkets in the semiarid region of Northeastern Brazil.Materials, Methods & Results: The study was carried out at commercial fairs in the state of Rio Grande do Norte, Braziliansemiarid region. The minimum number of eggs to be used was determined by the formula for simple random samples considering the following parameters: expected frequency of positive eggs of 50% (for sample maximization), 95% confidencelevel and sampling error of 10%. The minimum number of eggs to be sampled was 97; however, 128 eggs were obtained inthe period from August 2018 to April 2019. The internal content of the eggs was subjected to bacteriological culture usingan adapted methodology, and for the in vitro susceptibility assessment the disk diffusion method on Müller-Hintonagarwasused. In addition to biochemical tests, Gram-negative bacteria were subjected to microbiological diagnosis using theMALDI-TOF (Matrix Associated Laser Desorption-Ionization - Time of Flight) technique. Bacterial growth was found in40 (31.3%) eggs. The microorganisms most frequently isolated were Staphylococcus spp. (27.5%), Bacillus spp. (15%) andEnterobacter cloacae (25%). There was bacterial growth in albumens [n = 10; 7.8%] and in egg yolks [n=38; 29.7%] (P< 0.001). The antimicrobials that showed highest resistance rates were ampicillin...

Bacteria isolated from free-range chicken (Gallus gallus domesticus) eggs sold in semiarid conditions and its one health impact

Background: Food contamination is an important and growing public health concern due to the risk of foodborne illnesses.In this context, the egg, consumed all over the world, stands out. This food has been pointed out as a carrier of severalbacteria, causing outbreaks of food toxinfection. The production of free-range chicken (Gallus gallus domesticus) eggs hasbeen an alternative for generating income for producers in Brazil; however, there is no monitoring of the sanitary qualityof this product. The objective of this study was to identify microorganisms in 128 free-range chicken eggs sold in openmarkets in the semiarid region of Northeastern Brazil.Materials, Methods & Results: The study was carried out at commercial fairs in the state of Rio Grande do Norte, Braziliansemiarid region. The minimum number of eggs to be used was determined by the formula for simple random samples considering the following parameters: expected frequency of positive eggs of 50% (for sample maximization), 95% confidencelevel and sampling error of 10%. The minimum number of eggs to be sampled was 97; however, 128 eggs were obtained inthe period from August 2018 to April 2019. The internal content of the eggs was subjected to bacteriological culture usingan adapted methodology, and for the in vitro susceptibility assessment the disk diffusion method on Müller-Hintonagarwasused. In addition to biochemical tests, Gram-negative bacteria were subjected to microbiological diagnosis using theMALDI-TOF (Matrix Associated Laser Desorption-Ionization - Time of Flight) technique. Bacterial growth was found in40 (31.3%) eggs. The microorganisms most frequently isolated were Staphylococcus spp. (27.5%), Bacillus spp. (15%) andEnterobacter cloacae (25%). There was bacterial growth in albumens [n = 10; 7.8%] and in egg yolks [n=38; 29.7%] (P< 0.001). The antimicrobials that showed highest resistance rates were ampicillin...(AU)

Genital infection by Aerococcus viridans in a captive african elephant (Loxodonta africana) / Infecção genital por Aerococcus viridans em um elefante africano em cativeiro (Loxodonta africana)

Aerococcus viridans is an emerging pathogen for humans and livestock animals, mainly associated with genitourinary infections cases. Its occurrence in wild mammals has never been reported. The aim of this study was to determine the etiological agent associated with clinical a case of a genital infection in a female African elephant (Loxodonta africana). Phylogenetic analysis and antimicrobial susceptibility profile of the isolate were also addressed. The animal presented frequent cases of genital infection with intermittent white secretion. Purulent secretion was sampled and submitted to bacteriological exam. The isolate obtained was thus identified by phenotypic and molecular methods as A. viridans and was found to be similar to human pathogenic isolates in BLASTn and phylogenetic analysis. The isolate was sensitive to almost all antimicrobials evaluated, presenting resistance to ciprofloxacin and norfloxacin. This is the first report of occurrence of A. viridans infection in the genital tract of an African elephant.(AU)

Aerococcus viridans é um patógeno emergente para seres humanos e animais de produção, principalmente associado a casos de infecções geniturinárias. Sua ocorrência em mamíferos selvagens nunca foi relatada. O objetivo deste estudo foi determinar o agente etiológico associado a um caso clínico de infecção genital em uma fêmea de elefante africano (Loxodonta africana). Análises filogenéticas e perfil de susceptibilidade antimicrobiana do isolado também foram avaliados. O animal apresentou casos frequentes de infecção genital com eliminação de secreção branca intermitente. A secreção purulenta foi coletada e submetida a exame bacteriológico. O isolado obtido foi identificado por métodos fenotípicos e moleculares como A. viridans e apresentou alta similaridade a isolados humanos patogênicos nas análises de BLASTn e filogenética. O isolado foi sensível a quase todos os antimicrobianos avaliados, apresentando resistência à ciprofloxacina e norfloxacina. Este é o primeiro relato de ocorrência de infecção por A. viridans no trato genital de elefante africano.(AU)

Effects of physical and chemical factors on Ephemeroptera (Insecta) assemblages in an urban river of the eastern Colombian Llanos

The mayflies are insects widely known as indicators of aquatic quality in freshwater systems, they are abundant and diverse in tropical streams. In this study, it was determined the influence of physical, chemical and bacteriological characteristics on the Ephemeroptera assembly in a period of low precipitation in the Ocoa river of Meta department. There were 5,332 nymphs belonging to 4 families, 10 genera and 3 species. Three new genera records and one species are presented for Meta: Cloeodes, Zelusia, Lumahyphes and Americabaetis alphus. The genera Camelobaetidius and Varipes were found associated with conditions of contamination by organic matter and water mineralization. Likewise, Americabaetis, Thraulodes, Lumahyphesand Tricorythodes were associated with contamination conditions, especially with nitrites and nitrates. Nanomis, Cloeodes and Zelusia showed greater sensitivity to the contamination condition and were related to high percentages of dissolved oxygen in the water and they were in the sampling stations of the upper part in the river. It is important to mention that there were not Ephemeroptera in most of the stations associated with the urban area. That reflects their high sensitivity to polluting conditions at low precipitation period.

Effects of physical and chemical factors on Ephemeroptera (Insecta) assemblages in an urban river of the eastern Colombian Llanos

The mayflies are insects widely known as indicators of aquatic quality in freshwater systems, they are abundant and diverse in tropical streams. In this study, it was determined the influence of physical, chemical and bacteriological characteristics on the Ephemeroptera assembly in a period of low precipitation in the Ocoa river of Meta department. There were 5,332 nymphs belonging to 4 families, 10 genera and 3 species. Three new genera records and one species are presented for Meta: Cloeodes, Zelusia, Lumahyphes and Americabaetis alphus. The genera Camelobaetidius and Varipes were found associated with conditions of contamination by organic matter and water mineralization. Likewise, Americabaetis, Thraulodes, Lumahyphesand Tricorythodes were associated with contamination conditions, especially with nitrites and nitrates. Nanomis, Cloeodes and Zelusia showed greater sensitivity to the contamination condition and were related to high percentages of dissolved oxygen in the water and they were in the sampling stations of the upper part in the river. It is important to mention that there were not Ephemeroptera in most of the stations associated with the urban area. That reflects their high sensitivity to polluting conditions at low precipitation period.(AU)