Artigos - Produção Animal - FMVZ

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Zinc methionine or zinc sulphate supplementation modulate the development of the hypopharyngeal gland and expression of major royal jelly protein genes in Apis mellifera L. bees
(2023-01-01) do Prado Ribeiro, Giovanna [UNESP]; Kadri, Samir Moura [UNESP]; Justulin, Luis Antônio [UNESP]; Ribolla, Paulo Eduardo Martins [UNESP]; Orsi, Ricardo de Oliveira [UNESP]; Universidade Estadual Paulista (UNESP)
We analysed whether the source of zinc (inorganic and organic) influences the morphology of the hypopharyngeal gland (HPG) and the relative expression of major royal jelly protein (MRJP) genes in nursing bees at 6 days of age. For this, 15 colonies of Apis mellifera bees were subjected to the following treatments: Zn0 (control), zinc methionine (50 ppm) and zinc sulphate monohydrate (50 ppm) during the off-season (characterized by food reduction). MRJP1 and MRJP5 gene expression was analysed using real-time PCR and HPG sections under a microscope. Bee mortality and population development were also evaluated for 30 days. It was observed that the honeybees consumed all sugar syrup containing zinc both in its organic and inorganic forms, which did not promote mortality or affect population development. Regarding HPG morphometry, it was observed that zinc, regardless of the source, promoted the acini area significantly greater than that of the control. Zn methionine promoted a significant increase in HPG area compared with that of an inorganic source. The relative expression of MRJP1 and MRJP5 was not significantly affected compared with the control. We conclude that the mineral Zn promotes an increase in the HPG area and does not alter the pattern of expression of the MRJP1 and MRJP5 genes, and that the organic source of the mineral presents the best results.
Image feature extraction via local binary patterns for marbling score classification in beef cattle using tree-based algorithms
(2023-01-01) Pinto, Diógenes Lodi; Selli, Alana; Tulpan, Dan; Andrietta, Lucas Tassoni; Garbossa, Pollyana Leite Matioli; Voort, Gordon Vander; Munro, Jasper; McMorris, Mike; Alves, Anderson Antonio Carvalho; Carvalheiro, Roberto [UNESP]; Poleti, Mirele Daiana; Balieiro, Júlio Cesar de Carvalho; Ventura, Ricardo Vieira; Universidade de São Paulo (USP); University of Guelph; AgSights; Livestock Research Innovation Corporation; University of Wisconsin-Madison; Universidade Estadual Paulista (UNESP)
The objective of this study was to investigate the potential of creating a pipeline to classify the marbling score obtained from ribeye area (REA) images using computer vision and machine learning methods. Our database consisted of images and measurements (N = 2,446) from the transversal cut between the 12th and 13th ribs of the Longissimus dorsi muscle from carcasses of a beef cattle population (Bos taurus). Each sample was previously labeled by the industry using a low, medium or high marbling score. The prediction accuracies of two tree-based Machine Learning (ML) algorithms (Decision Tree - DT and Random Forest - RF) were compared, as well as different measures for extracting features from the REA images, which were used as input for the ML algorithms. In order to extract features based on detectable color patterns and textures contained in smaller parts of the grayscale image, we proposed the application of the local binary pattern (LBP) method prior to the adoption of ML methods. Mean classification accuracies for the test set ranged from 45.78% to 91.25% for different test scenarios. The results were mostly impacted by the feature extraction metrics, ML methods, potential subjectivity during the classification process by the industry, and the number of classes evaluated together. The best prediction accuracy results were achieved after performing the cross-validation (20% in each balanced group, 5 folds, and 10 repetitions), considering solely the extreme groups (low and high marbling scores) and pre-selecting from each group the 400 most visually representative samples. The RF algorithm outperformed the DT for most scenarios. After increasing the number of images to 580 samples for the same two groups, the highest testing accuracies were reduced to 83.05% for RF and 75.58% for DT. Such a decrease in the classification accuracies may be associated with the addition of erroneously classified images, due to the subjective nature of the industry evaluation. In conclusion, our preliminary studies showed the LBP method as a powerful feature extraction strategy considering a scenario where the labels were well defined. Our results revealed high accuracies for the classification of marbling extremes, but there is an evident need to improve the understanding of the biological and visual aspects that led to a sharp drop in classification accuracy after the insertion of the intermediate groups of marbling. In addition, the authors highlight the importance of an accurate labeling process for achieving better classification accuracy when applying supervised classification methods.
Field relevant doses of the fipronil affects gene expression in honey bees Apis mellifera
(2022-12-01) Astolfi, Aline [UNESP]; Kadri, Samir Moura [UNESP]; de Castro Lippi, Isabella Cristina [UNESP]; Mendes, Daniel Diego [UNESP]; Alonso, Diego Peres [UNESP]; Ribolla, Paulo Eduardo Martins [UNESP]; de Oliveira Orsi, Ricardo [UNESP]; Universidade Estadual Paulista (UNESP); Departamento de Produção Animal e Medicina Veterinária Preventiva
We analyzed the changes in the gene expression of forage honey bees (Apis mellifera) exposed to fipronil field-relevant dose (2.5 ppb/bee) by 1 or 4 h. A reduction in the expression of nine genes was observed after 4 h of exposure, five of which are related to the digestive system: endoglucanase E-4, MSS11 transcription activator, inositol monophosphatase 2-X1 transcription variant, ATP-binding cassette G subfamily, and cuticular protein 28; one related to exoskeleton composition: cuticular protein 28; two related to vitamin E transport and antioxidant system: alpha-tocopherol transfer protein-like and transcript variant X3. LOC551765, LOC100578929, and LOC102656070 were downregulated; however, these genes have not yet been studied. The results indicate that the fipronil causes changes in the expression of genes related to physiological and morphological, metabolism, and behavior in A. mellifera honey bees.
Effect of yerba mate (Ilex paraguariensis) in lamb diets on fatty acid profile, physical and sensory characteristics of the Longissimus muscle
(2022-11-01) Pena-Bermudez, Yuli A.; Lobo, Richard Roberto; de Amorim, Tamyres Rodrigues; Rojas-Moreno, Danny A.; Rodriguez-Aguilar, Daniel; Poleti, Mirele Daiana; Pereira, Angélica Simone Cravo; Pinheiro, Rafael Silvio Bonilha [UNESP]; Claudio da silva Bueno, Ives; Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP); University of Florida; National University of Colombia
This experiment studied the effects of yerba mate extract (YME) on lamb feed as a natural antioxidant in the fatty acid profile, physical and sensory traits of the meat of 36 lambs fed for 53 days. The treatments followed the randomized block design, with a control group without YME (0%) and three groups with 1, 2, and 4% of YME inclusion in the dry matter (DM). The inclusion of YME did not significantly affect the quality and sensory characteristics of the meat. However, it has improved the value of b* (p = 0.01) for the treatment with 4% YME, as well as the branched-chain fatty acids (BCFA), exhibited a diet effect (p = 0.01) for the same group. The addition of 4% of YME improves the composition and general acceptance of lamb meat, recommended to use it in animal feed.
Providing environmental enrichments can reduce subclinical spondylolisthesis prevalence without affecting performance in broiler chickens
(2023-04-01) Da Silva, Marconi Italo Lourenço [UNESP]; De Lima Almeida Paz, Ibiara Correia [UNESP]; Jacinto, Andressa Silva [UNESP]; Filho, Marcos Antonio Nascimento [UNESP]; De Oliveira, Ana Beatriz Santos [UNESP]; Dos Santos, Ingrid Grazieli Althman [UNESP]; Dos Santos Mota, Francine [UNESP]; Caldara, Fabiana Ribeiro; Jacobs, Leonie; Universidade Estadual Paulista (UNESP); Virginia Tech; Federal University of Grande Dourados
Environmental enrichment can increase the occurrence of natural behavior and improve leg health and other animal welfare outcomes in broiler chickens. This study aimed to assess the effects of three environmental enrichments, specifically hay bales, step platforms, and laser lights, on subclinical spondylolisthesis prevalence, productivity, behavior, and gait of broiler chickens (Gallus gallus domesticus). Twenty-four hundred day-old male Ross® AP95 chicks from a commercial hatchery were used in a completely randomized design with four treatments and four replicate pens per treatment. Pens contained either a Control (C) treatment, an environment similar to a commercial broiler chicken system without environmental enrichments, or an environment with either additional hay bales (HB), additional step platforms (SP), or additional laser lights (LL). Performance, yield, behavior (frequencies), gait score, and subclinical spondylolisthesis prevalences were assessed. When raised with SP or LL access, fewer chickens had subclinical spondylolisthesis than chickens without enrichments (C) or with HB access. Chickens with access to SP exhibited higher wing yield and less abdominal fat than animals from the C group. Chickens from the LL and HB treatments explored more and rested less frequently than animals from the C and SP treatments. As chickens aged, they became less active, exploring less and increasing resting and comfort behaviors. Treatments did not affect gait. Gait was not associated with subclinical spondylolisthesis prevalence. Environmental enrichments benefitted chicken health (subclinical spondylolisthesis) and behavior (exploration) without negative consequences for performance and yield. Copyright:
Molecular identification and antimicrobial resistance pattern of Nocardia isolated from 14 diseased dogs and cats
(2023-06-01) Condas, Larissa Anuska Zeni [UNESP]; de Farias, Marconi Rodrigues; Siqueira, Amanda Keller [UNESP]; Salerno, Tatiana [UNESP]; Chi, Kung Darh; Werner, Juliana; de Vargas, Agueda Castagna; Bond, Guilherme Borges; Gonoi, Tohru; Matsuzawa, Tetsuhiro; Ribeiro, Márcio Garcia [UNESP]; UniBrasil Centro Universitário; Universidade Estadual Paulista (UNESP); Pontifícia Universidade Católica do Paraná; Laboratório de Histopatologia Veterinária Werner & Werner; Universidade Federal de Santa Maria; Chiba University; University of Nagasaki
Nocardia are ubiquitous, saprophytic and opportunistic bacteria. They cause a set of pyogenic clinical infections in animals and humans, particularly immunocompromised patients, mostly affecting the skin and respiratory tract, with refractoriness to conventional therapy. The most descriptions of nocardial infections in companion animals involve case reports, and there are scarce case series studies focused on canine and feline nocardiosis in which diagnosis has been based on molecular techniques. We investigated epidemiological aspects, clinical findings, in vitro susceptibility profile, and molecular identification of Nocardia using PCR-based method targeted 16S rRNA gene in twelve dogs and two cats. Among dogs were observed cutaneous lesions (8/12 = 67%), pneumonia (3/12 = 25%), and encephalitis (2/12 = 17%), whereas cats developed cutaneous lesions and osteomyelitis. Nocardia and canine morbillivirus coinfection was described in six dogs (6/12 = 50%). A high mortality rate (6/8 = 75%) was seen among dogs. Three dogs (3/4 = 75%) and one cat (1/2 = 50%) with systemic signs (pneumonia, encephalitis, osteomyelitis), and 83% (5/6) of dogs with a history of concomitant morbillivirus infection died. N. nova (5/12 = 42%), N. cyriacigeorgica (3/12 = 25%), N. farcinica (2/12 = 17%), N. veterana (1/12 = 8%), and N. asteroides (1/12 = 8%) species were identified in dogs, whereas N. africana and N. veterana in cats. Among the isolates from dogs, cefuroxime (12/12 = 100%), amikacin (10/12 = 83%), gentamycin (10/12 = 83%), and imipenem (10/12 = 83%) were the most effective antimicrobials, whereas cefuroxime, cephalexin, amoxicillin/clavulanic acid, imipenem, and gentamycin were efficient against isolates from cats. Multidrug resistance was observed in 36% (5/14) of isolates. We describe a variety of Nocardia species infecting dogs and cats, multidrug-resistant ones, and a high mortality rate, highlighting a poor prognosis of nocardiosis in companion animals, particularly among animals systemically compromised or coinfected by canine morbillivirus. Our study contributes to species identification, in vitro antimicrobial susceptibility profile, clinical-epidemiological aspects, and outcome of natural Nocardia-acquired infections in dogs and cats.
Performance and Welfare of Sows Exposed to Auditory Environmental Enrichment in Mixed or Collective Housing Systems
(2023-04-01) Palermo Mendes, Janaina; Ribeiro Caldara, Fabiana; de Castro Burbarelli, Maria Fernanda; Valentim, Jean Kaique; Ferreira de Brito Mandú, Daniela; Garófallo Garcia, Rodrigo; Correia de Lima Almeida-Paz, Ibiara [UNESP]; Markiy Odakura, Agnes; Lourenço da Silva, Marconi Italo [UNESP]; Federal University of Grande Dourados (UFGD); Universidade Estadual Paulista (UNESP)
The research aimed to assess the effects of auditory environmental enrichment on sows in mixed housing (caged until 35 days after insemination and then collective pens) or collective housing (caged until 72 h after insemination and then collective pens). Reproductive performance, body surface and eye temperature (ET) were evaluated as sows’ welfare indicators. A sample of 56 sows between 2nd and 6th parity was submitted to the treatments from artificial insemination to weaning. The sows were assigned in a randomized block design with a 2 × 2 factorial scheme of treatments: mixed housing—control (MH-C), collective housing—control (CH-C), mixed housing—music (MH-M) and collective housing—music (CH-M). Auditory enrichment consisted of exposing sows daily throughout gestation and lactation to 6 h of classical music divided into 2 h periods. The ET of pregnant sows in collective housing was lower than that of sows in mixed housing (33.77 °C vs. 34.91 °C). Pregnant and lactating sows exposed to auditory environmental enrichment exhibited lower ET compared to those that had no access to the stimulus (pregnant 33.36 °C vs. 34.32 °C and lactating 34.21 °C vs. 34.83 °C). No housing type effect was found on the reproductive performance parameter; however, piglets from sows submitted to auditory environmental enrichment, regardless of the type of housing during gestation, were heavier at weaning (6.32 kg vs. 5.57 kg). Collective or mixed housing does not affect sows’ reproductive performance; perhaps, auditory environmental enrichment reduced stress in the gestation and lactation phases and provided greater piglet weight gain at weaning.
Oxidative stress in plants and the biochemical response mechanisms
(2023-01-01) Borges, Cristine Vanz [UNESP]; Orsi, Ricardo Oliveira [UNESP]; Maraschin, Marcelo; Lima, Giuseppina Pace Pereira [UNESP]; Universidade Estadual Paulista (UNESP); Universidade Federal de Santa Catarina (UFSC)
The accumulation of reactive oxygen species (ROS) caused by abiotic or biotic stresses generates oxidative stress in plant cells. To prevent the spread or to stop this chain reaction, antioxidant systems are activated, transforming free radicals (oxidants) into less toxic molecules for the cells. These generated antioxidant compounds can reduce the harmful effects of ROS and are classified as enzymatic (superoxide dismutase, peroxidases, catalase, among others) and nonenzymatic (bioactive amines, phenolic and carotenoid compounds). These molecules are essential to avoid damage to cell structures and cell death. In this chapter, we will present the response of plants under stress, activating some enzymatic systems and bioactive compounds (nonenzymatic), in order to alleviate oxidative stress.
Providing environmental enrichments can reduce subclinical spondylolisthesis prevalence without affecting performance in broiler chickens
(2023-01-01) Lourenço da Silva, Marconi Italo [UNESP]; Almeida Paz, Ibiara Correia de Lima [UNESP]; Jacinto, Andressa Silva [UNESP]; Nascimento Filho, Marcos Antonio [UNESP]; Oliveira, Ana Beatriz Santos de [UNESP]; Santos, Ingrid Grazieli Althman Dos [UNESP]; Mota, Francine Dos Santos [UNESP]; Caldara, Fabiana Ribeiro; Jacobs, Leonie; Universidade Estadual Paulista (UNESP); Virginia Tech; Federal University of Grande Dourados
Environmental enrichment can increase the occurrence of natural behavior and improve leg health and other animal welfare outcomes in broiler chickens. This study aimed to assess the effects of three environmental enrichments, specifically hay bales, step platforms, and laser lights, on subclinical spondylolisthesis prevalence, productivity, behavior, and gait of broiler chickens (Gallus gallus domesticus). Twenty-four hundred day-old male Ross® AP95 chicks from a commercial hatchery were used in a completely randomized design with four treatments and four replicate pens per treatment. Pens contained either a Control (C) treatment, an environment similar to a commercial broiler chicken system without environmental enrichments, or an environment with either additional hay bales (HB), additional step platforms (SP), or additional laser lights (LL). Performance, yield, behavior (frequencies), gait score, and subclinical spondylolisthesis prevalences were assessed. When raised with SP or LL access, fewer chickens had subclinical spondylolisthesis than chickens without enrichments (C) or with HB access. Chickens with access to SP exhibited higher wing yield and less abdominal fat than animals from the C group. Chickens from the LL and HB treatments explored more and rested less frequently than animals from the C and SP treatments. As chickens aged, they became less active, exploring less and increasing resting and comfort behaviors. Treatments did not affect gait. Gait was not associated with subclinical spondylolisthesis prevalence. Environmental enrichments benefitted chicken health (subclinical spondylolisthesis) and behavior (exploration) without negative consequences for performance and yield.
Flies (Insecta, Diptera) collected in the environment of dairy farms as carriers of Rotavirus A and betacoronavirus
(2023-03-01) Bertolini, Amanda Bezerra [UNESP]; Thyssen, Patrícia Jacqueline; Brandão, Paulo Eduardo; Prado, Aline Marrara; Silva, Sheila Oliveira de Souza; Mioni, Mateus de Souza Ribeiro [UNESP]; de Gouvea, Fábio Lucas Rezende [UNESP]; Pantoja, José Carlos de Figueiredo [UNESP]; Langoni, Helio [UNESP]; Guimarães, Felipe Freitas [UNESP]; Joaquim, Sâmea Fernandes [UNESP]; Guerra, Simony Trevizan [UNESP]; Leite, Domingos da Silva; Rall, Vera Mores [UNESP]; Hernandes, Rodrigo Tavanelli [UNESP]; Lucheis, Simone Baldini [UNESP]; Rossi, Gabriel Augusto Marques; Ribeiro, Márcio Garcia [UNESP]; Universidade Estadual Paulista (UNESP); Universidade Estadual de Campinas (UNICAMP); Universidade de São Paulo (USP)
AIMS: We aimed to investigate the prevalence of rotavirus and coronavirus in dipterans that commonly inhabit the environment of dairy farms. METHODS AND RESULTS: We collected 217 insect specimens from nine dairy farms, which were examined through hemi-nested RT-PCR followed by Sanger sequencing in search of VP1 and N genes for rotavirus and bovine coronavirus-BCoV, respectively. With a predominance of Muscidae (152/217 = 70%) 11 families of Diptera were identified. Rotavirus A (RVA) and betacoronavirus (BCoV) were detected in 14.7% (32/217) and 4.6% (10/217) of the dipterans, respectively. Sequencing of the amplicons was possible for 11.5% (25/217) of RVA and 0.5% (1/217) of BCoV, confirming the presence of these pathogens. CONCLUSIONS: Our findings highlight the role of dipterans as carriers of RVA and BCoV of great relevance for public and animal health.
Cutaneous protothecosis in a dog successfully treated with oral itraconazole in pulse dosing
(2023-12-01) Gmyterco, Vanessa Cunningham; Jagielski, Tomasz; Baldasso, Gustavo; Bacher, Louise Helene; Ribeiro, Márcio Garcia [UNESP]; de Farias, Marconi Rodrigues; Pontifícia Universidade Católica do Paraná; University of Warsaw; Universidade Estadual Paulista (UNESP)
Background: Protothecosis is a rare infectious disease caused by unicellular, achlorophyllous, microalgae of the genus Prototheca, ubiquitously distributed in nature. The algae are emerging pathogens, whose incidence is increasing in both human and animal populations and serious systemic infections related to this pathogen have been increasingly described in humans in recent years. After mastitis in dairy cows, canine protothecosis is the second most prevalent form of the protothecal disease in animals. Here, we report the first case of chronic cutaneous protothecosis due to P. wickerhamii in a dog in Brazil, successfully treated with a long-term therapy with itraconazole in pulse. Case presentation: Upon clinical examination, exudative nasolabial plaque, ulcered, and painful lesions in central and digital pads and lymphadenitis were observed in a 2-year-old mixed-breed dog, with a 4-month history of cutaneous lesions and contact with sewage water. Histopathological examination revealed intense inflammatory reaction, with numerous spherical to oval, encapsulated structures stained with Periodic Acid Schiff, compatible with Prototheca morphology. Tissue culture on Sabouraud agar revealed yeast-like, greyish-white colonies after 48 h of incubation. The isolate was subjected to mass spectrometry profiling and PCR-sequencing of the mitochondrial cytochrome b (CYTB) gene marker, leading to identification of the pathogen as P. wickerhamii. The dog was initially treated with oral itraconazole at a dosage of 10 mg/kg once daily. After six months, the lesions resolved completely, yet recurred shortly after cessation of therapy. The dog was then treated with terbinafine at a dose of 30 mg/kg, once daily for 3 months, with no success. The resolution of clinical signs, with no recurrence over a 36-months follow-up period, was achieved after 3 months of treatment with itraconazole (20 mg/kg) in pulse intermittently on two consecutive days a week. Conclusions: This report highlights the refractoriness of skin infections by Prototheca wickerhamii with therapies proposed in the literature and suggests a new treatment option with oral itraconazole in pulse dosing for long-term disease control successfully performed in a dog with skin lesions.
Colony Transport Affects the Expression of Some Genes Related to the Apis mellifera l. Immune System
(2022-12-01) Scaloppi, Maurice F. [UNESP]; Kadri, Samir M. [UNESP]; Mendes, Daniel D. [UNESP]; Ribolla, Paulo Eduardo M. [UNESP]; Orsi, Ricardo O. [UNESP]; Universidade Estadual Paulista (UNESP)
Migratory beekeeping can harm the bee colonies if not executed properly. Here, colonies of Apis mellifera were transported (for one or two hours) or not, following proper technical standards. To analyze gene expression (defensin-1, abaecin, and HSP70), forager bees were collected immediately, 24, and 72 hours after transportation. Bee mortality and population growth were measured before and after transportation. This study concludes that transporting honey bee colonies for 2 hours promotes immune system gene expression, although there are no significant changes in bee mortality and population growth of the colonies.
Evaluation of probiotic and glutamine in ovo on broilers challenged with Salmonella Heidelberg
(2023-03-01) de Souza Oro, Camila [UNESP]; dos Ouros, Caio Cesar [UNESP]; Ribeiro, Gabriella Costa [UNESP]; Tertuliano dos Santos, Cauê Bastos [UNESP]; Okamoto, Adriano Sakai [UNESP]; Hataka, Alessandre [UNESP]; de Moraes, Gustavo Nunes [UNESP]; Milbradt, Elisane Lenita [UNESP]; Filho, Raphael Lúcio Andreatti [UNESP]; Universidade Estadual Paulista (UNESP)
Salmonella Heidelberg (SH) is a bacterium that causes great economic damage to Brazilian poultry production. The indiscriminate use of antibiotics in poultry farming induces resistance to this microorganism. In this way, natural products are alternatives for replacing antibiotics, also contributing to the maintenance of the intestinal health of birds. Thus, the objective of the experiment was to evaluate broiler chicken hatchability and anti-Salmonella effects of in ovo treatment with a Lactobacillus reuteri probiotic consortium and glutamine. The experimental design was completely randomized with seven treatments. The in ovo treatments evaluated (control, glutamine, probiotic, probiotic plus glutamine, and ceftiofur hydrochloride, and ceftiofur hydrochloride plus glutamine) were mixed with the vaccine against Marek's disease and infectious bursal disease. The exposure to SH occurred 24 h after chick placement. Data was subjected to analysis of variance, Tukey, and Dunn test, with a significance level of 5%. All treatments that received glutamine in ovo showed better (P < 0.05) hatchability. The treatment with the probiotic, with or without glutamine, showed a lower (P < 0.05) amount of SH when compared to the other treatments. Three days after exposure, the chicks that received the probiotic showed greater (P < 0.05) villus height and greater crypt depth in the duodenum, jejunum, and ileum compared to the chicks that received ceftiofur hydrochloride. Therefore, glutamine in ovo improved hatchability, while the probiotic in ovo reduced the amount of Salmonella Heidelberg, in addition to increasing the villus height and crypt depth in broilers.
Genome-Based Characterization of Multidrug-Resistant Escherichia coli Isolated from Clinical Bovine Mastitis
(2023-03-01) dos Santos Alves, Taila; Rosa, Vinícius Sanches; da Silva Leite, Domingos; Guerra, Simony Trevizan [UNESP]; Joaquim, Sâmea Fernandes [UNESP]; Guimarães, Felipe Freitas [UNESP]; de Figueiredo Pantoja, José Carlos [UNESP]; Lucheis, Simoni Baldini [UNESP]; Rall, Vera Lúcia Mores [UNESP]; Hernandes, Rodrigo Tavanelli [UNESP]; Langoni, Helio [UNESP]; Ribeiro, Márcio Garcia [UNESP]; Universidade Estadual de Campinas (UNICAMP); Universidade Estadual Paulista (UNESP)
Mastitis occurrence in dairy cows is a broad topic that involves several sectors, from antimicrobial resistance and virulence of strains to economic implications and cattle management practices. Here, we assessed the molecular characterization (antimicrobial resistance determinants, virulence genes, sequences type, serotypes, and plasmid types) of 178 Escherichia coli strains isolated from milk samples from cows with clinical mastitis using a genome-based k-mers approach. Of these, 53 (29.8%) showed multidrug resistance by disc diffusion. We selected eight multidrug-resistant mastitis-associated E. coli for whole-genome sequencing and molecular characterization based on raw data using k-mers. We assessed antimicrobial resistance genes, virulence factors, serotypes, Multilocus Sequence Typing (MLST), and plasmid types. The most antimicrobial resistance gene found were blaTEM-1B (7/8), tetA (6/8), strA (6/8), strB (6/8), and qnrB19 (5/8). A total of 25 virulence factors were detected encoding adhesins, capsule, enzymes/proteins, increased serum survival, hemolysin, colicins, and iron uptake. These virulence factors were associated with Extraintestinal Pathogenic E. coli. Three pandemic clones were found: ST10, ST101, and ST69. Two E. coli were assigned in the O117 serogroup and one in the O8:H25 serotype. The most common plasmid groups were IncFII (7/8) and IncFIB (6/8). Our findings contribute to the knowledge of virulence mechanisms, epidemiological aspects, and antimicrobial resistance determinants of E. coli strains obtained from clinical mammary infections of cows.
Characterization of mammary pathogenic Escherichia coli reveals the diversity of Escherichia coli isolates associated with bovine clinical mastitis in Brazil
(2023-02-01) Orsi, Henrique [UNESP]; Guimarães, Felipe F. [UNESP]; Leite, Domingos S.; Guerra, Simony T. [UNESP]; Joaquim, Sâmea F. [UNESP]; Pantoja, Jose C.F. [UNESP]; Hernandes, Rodrigo T. [UNESP]; Lucheis, Simone B.; Ribeiro, Márcio G. [UNESP]; Langoni, Helio [UNESP]; Rall, Vera L.M. [UNESP]; Universidade Estadual Paulista (UNESP); Universidade Estadual de Campinas (UNICAMP); Paulista Agency of Agribusiness Technology
Mammary pathogenic Escherichia coli (MPEC) is one of the most common pathogens associated with clinical mastitis. We analyzed isolates obtained from milk samples of cows with clinical mastitis, collected from 10 farms in Brazil, to verify molecular and phenotypic characteristics. A total of 192 (4.5%) mammary pathogenic E. coli isolates were obtained from 4,275 milk samples analyzed, but we tested 161. We assigned most of these isolates to E. coli phylogroups B1 (52.8%) and A (36.6%), although phylogroups B2, C, D, E, and unknown also occurred. All isolates were assessed for the presence of several genes encoding virulence factors, such as adhesins (sfaDE, papC, afaBC III, ecpA, fimH, papA, and iha), toxins (hlyA, cnf1, sat, vat, and cdt), siderophores (iroN, irp2, iucD, ireA, and sitA), an invasion protein (ibeA), and serum resistance proteins (traT, KpsMTII, and ompT), and isolates from phylogroups B1, B2, and E showed up to 8 genes. Two isolates harbored the locus of enterocyte effacement (escN+) and lack the bundle-forming pilus (bfpB−) operon, which corresponds to a molecular profile of a subgroup of diarrheagenic E. coli (aEPEC), thus being classified as hybrid MPEC/aEPEC isolates. These isolates displayed a localized adherence-like pattern of adherence in HeLa cells and were able to promote F-actin polymerization underneath adherent bacteria. Based on the pulsed-field gel electrophoresis analyses, considerable genetic variability was observed. A low index of antimicrobial resistance was observed and 2 extended-spectrum β-lactamase–producing E. coli were identified, both harboring blaCTX-M15 gene, and were classified as ST10 and ST993 using multilocus sequence typing. A total of 148 (91.2%) isolates were weak biofilm producers or formed no biofilm. Because raw milk is still frequently consumed in Brazil, the occurrence of virulence factor–encoding genes from extraintestinal or diarrheagenic E. coli added to the presence of extended-spectrum β-lactamase–producing isolates can turn this veterinary medicine problem into a public health concern.
Virulence-encoding genes related to extraintestinal pathogenic E. coli and multidrug resistant pattern of strains isolated from neonatal calves with different severity scores of umbilical infections
(2023-01-01) Martins, Lorrayne de Souza A. [UNESP]; Motta, Rodrigo G.; Martinez, Antonio C.; Orsi, Henrique [UNESP]; Hernandes, Rodrigo T. [UNESP]; Rall, Vera L.M. [UNESP]; Pantoja, José C.F. [UNESP]; Nardi Júnior, Geraldo de; Ribeiro, Márcio G. [UNESP]; Universidade Estadual Paulista (UNESP); State University of Maringá; Fatec
Umbilical infections in calves comprise a major cause of neonatal mortality and have been related to a variety of microorganisms. E. coli is an opportunistic enteropathogen characterized by a diversity of virulence factors (VF). Nonetheless, the gene profiles that encode VF associated with umbilical infections in calves and their effect on the clinical severity remains unclear. In this scenario, microbial identification (with an emphasis on E. coli), was carried out among 150 neonatal calves (≤30 days of age) with umbilical infections, where the omphalopathies were clinically scored as mild, moderate, or severe. Also, a panel of 16 virulence-encoding genes related to extraintestinal pathogenic E. coli (ExPEC) were investigated, i.e., fimbriae/adhesins (sfa/focDEa, papA, papC, afaBC), toxins (hlyA, sat, cnf1, cdt), siderophores (iroN, irp2, iucD, ireA), invasins (ibeA), and serum resistance (ompT, traT, kpsMT II). Bacteria and yeasts isolates were identified using mass spectrometry. Bacteria, yeasts, and fungi were isolated in 94.7% (142/150) of neonatal calves sampled. E. coli was the agent most frequently isolated (59/150 = 39.3%), in pure culture (27/59 = 45.8%) and combined infections (32/59 = 54.2%), although a great variety (n = 83) of other species of microorganisms were identified. Clinical severity scores of 1, 2, and 3 were observed in 32.2% (19/59), 23.7% (14/59), and 44.1% (26/59) of E. coli infections, respectively. The ExPEC genes detected were related to serum resistance (traT, 42/59 = 72.2%; ompT, 35/59 = 59.3%, kpsMTII, 10/59 = 17%), invasins (ibeA, 11/59 = 18.6%), siderophores (iucD, 9/59 = 15.3%; iroN, 8/59 = 13.6%), and adhesins/fimbriae (papA, 8/59 = 13.6%; papC, 15/59 = 9.6%). The presence of each virulence gene was not associated with the case's clinical score. Among all isolates, 89.8% (53/59) showed in vitro resistance to sulfamethoxazole/trimethoprim and 59.3% to ampicillin (35/59), while 94.1% (55/59) revealed a multidrug resistant profile. Great complexity of bacteria, yeast, and fungi species was identified, reinforcing the umbilical infections of neonatal calves as a polymicrobial disorder. The high occurrence of E. coli (39.3%) highlights the role of this pathogen in the etiology of umbilical infections in calves. Furthermore, a panel of ExPEC genes was investigated for the first time among calves that were clinically scored for case severity. The high prevalence of traT and ompT indicates that these serum resistance-related genes could be used as biomarkers for further investigations of ExPEC isolates from umbilical infections. Our results contribute to the etiological investigation, clinical severity scoring, antimicrobial resistance pattern, and virulence-related to ExPEC genes involved in umbilical infections of neonatal calves.
Bacterial identification in cerebrospinal fluid of domestic species with neurologic signs: a retrospective case-series study in 136 animals (2005–2021)
(2023-03-01) Ribeiro, Márcio Garcia [UNESP]; Pereira, Thiago Tourinho [UNESP]; de Lima Paz, Patrik Júnior [UNESP]; de Almeida, Beatriz Oliveira [UNESP]; Cerviño, Carmen S. Araújo [UNESP]; Rodrigues, Carolina Aparecida [UNESP]; Santos, Gabrielly Terra Sartori [UNESP]; de Souza Freire, Larissa Maria [UNESP]; Portilho, Fábio Vinicius Ramos [UNESP]; Filho, Marcelo Fagali Árabe [UNESP]; Paschoal, Natália Rodrigues [UNESP]; Bello, Thaís Spessotto [UNESP]; Megid, Jane [UNESP]; Langoni, Helio [UNESP]; Appolinário, Camila Michele [UNESP]; Borges, Alexandre Secorun [UNESP]; Amorim, Rogério Martins [UNESP]; Giuffrida, Rogério; de Oliveira Filho, José Paes [UNESP]; Siqueira, Amanda Keller; Listoni, Fernando José Paganini [UNESP]; Paes, Antonio Carlos [UNESP]; Universidade Estadual Paulista (UNESP); University of Western São Paulo - UNOESTE; Midwestern State University - UNICENTRO
Central nervous system (CNS) infections comprise life-threatening clinical conditions in domestic species, and are commonly related to severe sequelae, disability, or high fatality rates. A set of bacterial pathogens have been identified in central nervous infections in livestock and companion animals, although the most of descriptions are restricted to case reports and a lack of comprehensive studies involving CNS-related bacterial infections have been focused on a great number of domestic species. In this scenario, we retrospectively investigated selected epidemiological data, clinical findings, bacteriological culture, and in vitro susceptibility patterns of 136 nonrepetitive neurologic cases in domestic species (2005–2021). Bacterial isolates were recovered from 25% (34/136) of the cerebrospinal fluid (CSF) sampled. The isolates were obtained from cattle (9/136 = 6.6%), dogs (7/136 = 5.1%), horses (6/136 = 4.4%), goats (3/136 = 2.2%), pigs (3/136 = 2.2%), sheep (3/136 = 2.2%), cats (2/136 = 1.5%), and asinine (1/136 = 0.7%). Among animals with bacterial isolation, Staphylococcus aureus (6/34 = 17.6%), Escherichia coli (5/34 = 14.7%), Staphylococcus beta-hemolytic (5/34 = 14.7%), and Trueperella pyogenes (3/34 = 8.8%) were predominant, in addition to a miscellaneous of other bacteria isolated in minor frequency, e.g., Corynebacterium pseudotuberculosis, Enterobacter cloacae, Mannheimia haemolytica, Pseudomonas aeruginosa, and Streptococcus equi subsp. equi. In vitro susceptibility tests of isolates revealed that amoxicillin/clavulanic acid (11/13 = 84.6%), cephalexin (9/11 = 81.8%), and florfenicol (9/12 = 75%) were the most effective antimicrobials. Conversely, isolates exhibited resistance mainly to tetracycline (6/10 = 60%), penicillin (6/11 = 54.5%), and trimethoprim/sulfamethoxazole (5/11 = 45.5%). Also, multidrug resistance to ≥ 3 classes of antimicrobials was found in 23.5% (8/34) strains. Data relative to the outcome was available in 79.4% (27/34) of animals that had bacterial isolation, and from these, the lethality rate was 92.6% (25/27). Incoordination (14/34 = 41.2%), recumbency (11/34 = 32.4%), apathy (10/34 = 29.4%), anorexia (9/34 = 26.5%), blindness (7/34 = 20.6%), seizure (6/34 = 17.6%), limb paresis (5/34 = 14.7%), head-pressing (4/34 = 11.8%), and nystagmus (3/34 = 8.8%) were the most frequent clinical signs. A variety of bacterial pathogens were identified in the CSF of domestic species showing neurologic signs, with a predominance of staphylococci, streptococci, and enterobacteria. High lethality of cases, poor in vitro efficacy of conventional antimicrobials, and a high in vitro multidrug resistance pattern of isolates were seen. Our results contribute to etiological characterization, antimicrobial resistance patterns, and clinical-epidemiological findings of bacterial infections in domestic species with neurological signs.
Advances in the crosstalk between maternal separation and voluntary ethanol consumption and effects on reproduction
(2022-12-15) Teixeira, Giovana Rampazzo [UNESP]; Martins, Otávio Augusto [UNESP]; Kremer, Rafael; Veras, Allice Santos Cruz [UNESP]; Pinheiro, Patricia Fernanda Felipe [UNESP]; Mello-Junior, Wilson [UNESP]; Martinez, Francisco Eduardo [UNESP]; Universidade Estadual Paulista (UNESP); Universidade Federal de Santa Catarina (UFSC)
Maternal separation (SM) is an event caused by early stress and may be associated with behavioral changes and vulnerabilities, enhancing ethanol consumption in adulthood. The aim of the study was to evaluate whether MS potentiates the effects of ethanol ingestion on physiological hormone regulation and its interference in testicular and epididymal morphofunctional aspects in voluntary ethanol-consuming rats. Therefore, for the first time, we investigated the effect of maternal separation and ethanol consumption in adulthood and for this we used free choice ethanol-consuming strains. Responses of metabolic and hormonal parameters were also addressed, as well as their effects on reproductive function. In summary, MS promoted an increase in voluntary ethanol consumption in UChA and UChB animals. There was an influence of MS on the increase of circulating corticosterone and testosterone in UChB animals (high-ethanol-preferring 10 % v/v). MS performed in the hyporesponsive period to stress promoted an increase in glucose and circulating lipids, as well as a reduction in lactate dehydrogenase levels. Daily sperm production and transit time through the epididymis in UChB animals were increased by MS. Together, these findings show that MS potentiates the effects of ethanol ingestion and promotes an imbalance in plasma hormone concentrations, interfering with the reproductive functional imbalance of ethanol-consuming rats.
Pregnancy Associated Glycoproteins Effects on Placenta Function, Uterine Remodeling and Attachment of Bovine Conceptuses.
(Oxford Univ Press Inc, 2022-10-01) Melo, Gabriela Dalmaso de; Franco, Gessica; Ortega, M. Sofia; Oliveira Filho, Ramiro V.; Bega, Amanda; Moraes Vasconcelos, Jose Luiz [UNESP]; Cerri, Ronaldo; Pohler, Ky G.; Texas A&M Univ; Univ Missouri; Univ British Columbia; Universidade Estadual Paulista (UNESP)
Atypical Mandibular Osteomyelitis in an Ewe Caused by Coinfection of Pseudomonas aeruginosa and Lactococcus raffinolactis
(Univ Fed Rio Grande Do Sul, 2022-01-01) Motta, Rodrigo Garcia; Martinez, Antonio Campanha; Marques Pereira Junior, Oduvaldo Camara; Araujo Martins, Lorrayne de Souza [UNESP]; Priscila da Paz, Jessica; Bello, Thais Spessotto [UNESP]; Arabe Filho, Marcelo Fagali [UNESP]; Ribeiro, Marcio Garcia [UNESP]; Universidade Estadual de Maringá (UEM); Universidade Estadual Paulista (UNESP)
Background: Osteomyelitis is defined as a bone inflammation involving the cortical and medullary regions, usually caused by the local invasion of opportunistic microorganisms. The inflammatory reaction of bone may extend to the periosteum and soft tissues, compromising adjacent structures far from the initially infected foci. Different classifications of transmission routes, gravity levels, and tissues involved in animal and human osteomyelitis are available. In humans, the infection can reach bone tissue by exogenous or hematogenous pathways. This paper reports an atypical case of mandibular pyogranulomatous osteomyelitis in an ewe caused by concomitant Pseudomonas aeruginosa and Lactococcus raffinolactis infection. Case: The animal presented a 1-month history of progressive mandibular enlargement refractory to conventional therapy. In a physical examination, an increased volume located in the ventrolateral region of the right ramus of the mandible was observed. Fine-needle aspiration of the lesion enabled isolation in bacteriological culture of Pseudomonas aeruginosa and Lactococcus raffinolactis using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Besides support care procedures and antimicrobial treatment approaches for the sheep based on in vitro tests, the animal died due to the severity of the clinical signs and the progressive worsening of the general health status. The radiographic image examination of the mandibular region revealed a severe and infiltrative periodontal reaction, with a predominance of a great number of neutrophils and macrophages, necrotic areas, and bone destruction, characterized histologically as a pyogranulomatous rection. At post mortem examination, a large pyogranuloma was observed in the entire horizontal branch of the mandible as well, showing a dark yellowish content of coarse consistency, caseous appearance, and bone fragmentation. Discussion: Ovine mandibular osteomyelitis is a well-established bone inflammation involving the cortical and medullary regions, characterized clinically by local enlargement, asymmetry, pain sensitivity, edema, hyperthermia, infiltrate caseous or suppurative material, and bone rarefaction. In the current report, 1-month history of progressive enlargement of the mandibular region, prostration, and weight loss in an ewe were referred. Where clinical and epidemiological features, bacteriological, cytological, histological, and mass spectrometry diagnostic approaches were assessed to diagnostic. Most reports involving the etiology of ovine mandibular osteomyelitis have been diagnosed based on classical phenotypic tests. Here, the concomitant identification of P. aeruginosa and L. raffinolactis infection was possible using mass spectrometry (MALDI-TOF), highlighting the importance of molecular methods in the diagnosis of animal diseases. In addition, the differentiation between Lactococcus and Enterococcus species is difficult, which could underestimate the diagnosis of Lactococcus species as a primary pathogen from animal diseases. We report, for the first time, a fatal case of mandibular pyogranulomatous osteomyelitis in a sheep caused by Pseudomonas aeruginosa and Lactococcus raffinolactis coinfection.

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