Identification of potentially harmful bacterial genera of veterinary relevance in the Llanquihue urban wetlands

Water bodies constitute natural, social, and ecological heritage under constant threat from the footprint of human action. They are the habitats of many species and play a key role in sustaining biodiversity in different ecosystems. However, anthropogenic activity can result in eutrophication of water bodies, favoring the excessive growth of microorganisms, which can be a risk factor for animal, human, and environmental health. The Agricultural and Livestock Service [Servicio Agrícola y Ganadero, (SAG)] of Chile developed a list of Mandatory Reportable Diseases (MRD) to report suspected contagious diseases of veterinary relevance. In this sense, the detection of microorganisms in water bodies for the characterization of their sanitary status is important for the development of monitoring strategies and the prevention of possible diseases. This study aimed to determine the presence of potentially harmful bacteria in Llanquihue urban wetlands. Here, we used metagenomic sequencing of amplicons to characterize the composition of microorganisms in three urban wetlands in the city of Llanquihue, which could be a risk factor for animal health. Our main finding was the detection of genera that may be associated with MRD, in addition to other microbial genera that have pathogenic potential. On the other hand, we also report the presence of cyanobacteria genera that can be involved in toxin production in water bodies as another potential source of risk. This is the first study to use massive sequencing techniques to analyze the sanitary status of urban wetlands in the city of Llanquihue.

Carvacrol-loaded invasomes efficacy against multidrug resistant isolates of Enterobacteriaceae and housefly

The current study aimed to evaluate the antimicrobial activity of carvacrol-loaded invasomes (CLI) against multidrug-resistant (MDR) Enterobacteriaceae and its mechanical vector, the housefly. CLI were prepared and characterized in the laboratory. Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica subsp. enterica serovar Enteritidis, Salmonella enterica subsp. enterica serovar Typhimurium, and Klebsiella oxytoca were among the MDR enterobacteriaceae stains investigated. These strains were first isolated and identified from naturally infected chickens. The antibacterial activity of CLI against the MDR isolates was evaluated using the diffusion method. In addition, the insecticidal activity of CLI against housefly larvae and pupae was tested. The MDR index of all evaluated isolates was greater than 20%, indicating that they were all multidrug-resistant. CLI decreased the growth of all isolates except S. Typhimurium and P. aeruginosa at a dose of 0.0125%; however, pure carvacrol inhibited the growth of only Klebsiella oxytoca. Furthermore, both CLI and pure carvacrol inhibited Klebsiella oxytoca growth at different concentrations. CLI inhibited E. coli and S. enteritidis at lower concentrations than pure carvacrol, even at a doubled concentration. Carvacrol and CLI caused significant larval mortality even at low concentrations, with LC50 reached at concentartions of 2.54 and 2.19 µl/ml, respectively. Furthermore, at a low concentration of 3.125 µl/ml, both elicited a high percentage inhibition rate (PIR) in pupae. In conclusion, CLI demonstrated substantial antibacterial action, particularly against MDR isolates, as well as pesticide activity against houseflies.

Experimental assessment of the efficacy of copper ion treatment against penicillin G contained in UHT milk and PBS

Antibiotics are widely used in animal production to treat bacterial infections and to improve performance and animal welfare. Their misuse poses a threat to public and animal health because of the possible development of antibiotic-resistant microorganisms. Among the many strategies that have been considered to address this problem are methods to degrade antibiotic residues, especially those from the human and animal food chain. This study describes the effect of copper ion treatment on the detection of penicillin G in a liquid matrix. An in vitro experimental study was designed using both commercial milk and PBS spiked with three different concentrations of penicillin G. Each sample was treated for 30 min with copper ions. All samples were tested for antibiotics before and after treatment using a commercial enzyme-linked receptor binding assay. Additionally, pH, copper concentration, and temperature were evaluated. Antibiotic residues were detected in all spiked PBS and milk samples before treatment with copper ions. However, after 30 min of treatment, no antibiotic residues were detected in any sample at any concentration tested. In conclusion, treatment of penicillin-contaminated milk and PBS samples with copper ions affects antibiotic detection, which would potentially reduce antibiotic levels

Decontamination protocols on bovine fecal and environmental samples for Mycobacterium avium ssp. paratuberculosis growth in solid media

Bacteriological culture of Mycobacterium avium subsp. paratuberculosis (MAP) is considered the gold standard to confirm its presence in several matrices for Johne’s disease diagnosis. Whether it is a liquid or solid culture, a problem with MAP culture is that non-interpretable results arise because of overgrowth by other microorganisms, making MAP growth and identification more difficult or impossible. We systematically reviewed published decontamination protocols and their effects on MAP culture from bovine fecal and environmental samples on solid media. Based on our findings, we suggest a step-by-step decontamination protocol. The OVID®/MEDLINE, PubMed®, SciELO Citation Index®, and Redalyc® platforms as well as the International Colloquium on Paratuberculosis (ICP) proceedings and the reference lists were reviewed to identify relevant studies. The inclusion criteria considered articles published in English, Portuguese, French, German, Spanish, and peer-reviewed journals. The exclusion criteria included unrelated topics, species other than bovines, other than environmental/faecal samples, other than diagnostic techniques of interest, and non-original articles. Definitive studies were obtained through the authors’ consensus regarding their eligibility and quality. In total, 1,004 publications matched the search terms, and 27 articles met the inclusion criteria, of which 45 derived and reported 15 different decontamination protocols. The centrifugation-one-step hexadecylpyridinium chloride protocol, which used over 22,154 fecal samples in three studies, was found to be the most suitable, reporting an average MAP isolation rate of 3.99% (886/22,154) and an average contamination rate of 0.17% (38/22,154). This systematic review highlights the need for further refinement of decontamination protocols to minimize the loss of viable MAP during processing of bovine fecal and environmental samples

Effects of selling through an auction market on physiological variables in beef calves

In Chile, selling cattle through auction markets is common, and involves various stressful events. The objective of this study was to determine the effects of selling weaned beef calves through an auction market on indicators of physiological stress. Considering that transportation is invariably linked to any commercialization procedure, we compared the effects of exposing calves to transport and sale through an auction market versus transporting them directly between farms. Twenty calves of 175 (+/-37.5) kg live weight were transported from their origin farm to an auction market (2.5 h journey), where 10 calves were unloaded and exposed to regular handling within the market (unloading, sorting, regrouping, penning, sale yard showing) and thereafter loaded and transported back to the farm (TM). The remaining 10 calves were not unloaded at the market and were immediately transported back to the farm, simulating a direct sale (TD). Data collection (blood samples, body temperature, and live weight) was performed at the farm in both groups before loading (initial, 0 h), at the end of the process, after unloading TM calves back on the farm (after unloading, 12 h), and during recovery at 24, 48, and 72 h after the initial sampling. We used mixed-effects models to compare treatments and samplings. No differences were found between treatments in terms of live weight loss, maximum eye temperature, plasma beta-hydroxybutyrate, and serum cortisol concentrations. The rectal temperature was higher (p < 0.05) in TM after treatment and remained higher (p > 0.05) in the samples than in TD. Calves that passed through the market showed higher (p < 0.05) plasma CK activity after unloading (12 h) than did those that did not. The mean glucose concentration increased (p < 0.05) at 12h and remained higher than the initial value (0 h) in both the calf groups. This was a preliminary study, and according to the results, experiments should be repeated to test more animals under different situations and premises

Selenium affects genes associated with immunity and apoptosis in in vitro follicles of ewes

Selenium plays an important role in bodily functions. It activates immune cells such as neutrophils and immunity through T cells. However, its role in ovarian follicular gene expression has not been reported. The effects of selenomethionine (SeMet) on gene expression in in vitro ewe follicles and their relationship with immunity and apoptosis were studied. Preovulatory follicles were randomly cultured, and SeMet was added to the culture, followed by incubation for 24 h. Total RNA was extracted from follicles and placed on a microarray chip. Gene expression was analyzed using GenArise and classified using the DAVID v6.8 bioinformatics program based on the Gene Ontology and Kyoto Encyclopedia for Genes and Genomes (KEGG). Gene pathways were designed using Cytoscape v3.7.2. There were 2,538 differentially expressed genes (DEGs): 1,228 upregulated and 1,310 downregulated. The upregulated genes were classified into 32, 18, and 12 nodes related to biological processes, cellular components, and molecular functions, respectively. KEGG assigned these genes to 41 metabolic and signaling pathways. 94 genes were involved in eight pathways associated with immune processes. The expression levels of CD8, NFAT2, and CD48 were quantified using RT-qPCR. Selenium activated gene expression in the in vitro preovulatory follicles of ewes, some of which were related to the immune system. The relationship among CD8, NFAT2, and CD48 genes suggests a possible immune pathway related to follicular apoptosis regulation

Frequency and risk factors of intestinal parasites in pet dogs from Mexicali, Mexico

Parasitic intestinal infections in dogs represent a problem for human health, because a wide variety of these parasites have zoonotic potential. Therefore, proximity to pets puts us at risk. The objective of this study was to determine the frequency and risk factors (age, sex, size, breed, presence of ectoparasites and gastrointestinal disorders) of intestinal parasites in the feces of dogs attending a Veterinary Hospital in the City of Mexicali, Baja California, Mexico. A total of 148 fecal samples were collected from canine patients and analyzed for parasite identification and parasite load. A 12.2% (18/148) of the samples were positive to parasitic intestinal infections. The frequency of specific infections was an 8.1% of Cryptosporidium sp., followed by a 2.7% of Cystoisospora sp., and 1.4% of Toxascaris leonina. A statistical significance was identified between the presence of intestinal parasites and mongrel breed. The predominance of protozoa shows the importance of diagnosis prior to treatment with anthelminthic drugs, since preventive antiparasitic protocols are commonly used, although these particular parasites are out of the spectrum of those drugs. Cryptosporidium spp. have zoonotic potential, particularly in immunocompromised patients, and there are few or no treatment options

From Domination to Dialogue and the Ethics of the Between:Transforming Human-Working Equine Relationships in Mountain Tourism

The welfare of working equines is born of the relationship co-created with humans and the relational practices humans bring to this work. Our understanding of this relationship remains elusive, however, for it involves attending to that which arises both within and between the equine and the human. Attempts to study such relationships have, arguably, been confounded by the liminalities of relational practices, power literacy and the limitations of language, propositional knowing and the dualistic thinking that characterises many scientific disciplines. This paper presents the theoretical framework that underpins an experiential awareness-based Action Research approach to transforming human-equine relations within the international mountain tourism industry. This approach privileges curiosity, compassion and primary or contemplative knowing and the development of self-awareness. Drawing on the work of Martin Buber on genuine dialogue and of Otto Scharmer on generative dialogue, this paper provides insights into what can arise in the between when attitudinal shifts are encouraged and facilitated that allow humans and equines to meet genuinely and be fully present to each other. This ultimately involves surrendering control, letting go, the dissolution of subject-object awareness and access to non-dualistic ways of knowing. An awareness of the importance of such shifts and of the source from which we operate is of fundamental importance to the realisation of the co-creative project that humans and equines can engage in. Failure to appreciate this distinction, arguably, leads and gives rise to relationships, whether human-to-human or human-to-horse, characterised by domination rather than partnering, absencing rather than presencing, by monologue rather than dialogue. The ethical and practical implications of this awareness are profound, with implications felt at the level of the individual, for whom the I deepens the more you pay attention, and at the level of the relationship, but also at the level of communities, whether these be constituted locally, nationally, internationally or indeed globally

Physical quality of different industrial versus non-industrial eggs obtained from groceries and markets in southern Chile

The aim of this study was to determine external and internal quality parameters of industrial (cages and cage-free) and family farms eggs that are normally available at groceries in developing countries such as Chile. Two experiments were performed to evaluate 1) quality differences between family farms and industrial eggs and 2) to determine quality differences between brown shell eggs from different industrial cage and cage-free systems. Experiment 1 consisted of five groups where three of them were industrial eggs: i) cage white shell eggs, ii) cage brown shell eggs, iii) brown shell cage-free eggs; and two of them were non-industrial: iv) family farm brown shell eggs and v) family farm blue shell eggs. Experiment 2 had four groups, all brown-shell types of eggs were used: i) cage brown eggs, ii) cage-free from aviary eggs, iii) southern free-range eggs and iv) central free-range eggs. In both Experiments, egg weight, egg length, egg width, egg shape index, Haugh units, albumen ratio, egg yolk, yolk weight and albumen weight, blood and meat spots were determined. In Experiment 1, brown and blue-shelled family farm eggs were equal in terms of external and internal quality, except for blood spots, with brown eggs having more incidence. In Experiment 2, free-range eggs presented more intense yolk colors compared to those from battery and cages. In both experiments, free-range eggs presented the darker yolk color. It can be concluded that brown and blue-shelled family farm eggs are equal in terms of external and internal quality, except for blood spots, with brown eggs having more incidence. In addition, free-range eggs from the southern part of the country presented better shell quality, whereas free-range eggs presented more intense yolk colors, while those of battery

Evolution of IFN-gamma response against mycobacterial antigens used for the diagnosis of bovine tuberculosis in BCG vaccinated cattle under a natural transmission setting in central Chile

Bovine tuberculosis (bTB) is a chronic disease of animals mainly caused by Mycobacterium bovis, a zoonotic pathogen that generates economic losses in the milk and meat industry. In central Chile, the Metropolitan Region concentrates dairy herds with the highest bTB prevalence of the country and the official veterinary service has supported the evaluation of the M. bovis Bacillus Calmette-Guerin (BCG) vaccine in this area with the replacement of tuberculin purified protein derivative (PPDs) by the DIVA (Differentiating Infected from Vaccinated Animals) peptides for the bTB diagnosis in the herds. This study aimed to describe the IFN-gamma response against PPDs (bovine and avian PPD) and DIVA antigenic cocktails (ESAT-6/CFP-10 and Rv3615c) in BCG vaccinated 11-month-old heifers under a natural transmission scenario. Sixty-two animals were vaccinated via subcutaneous route with a 2-8 x 105 colony forming units of BCG Russia strain and 60 control animals received sterile saline. Blood sampling was performed at time 0, previous to vaccination, and then at 3, 6, 9, 12, 15 and 18 months post-inoculation. The follow up of the IFN-gamma response in animals determined that the BCG vaccination interferes with the diagnosis of bTB using the traditional bovine PPD between 9 and 12 months post-inoculation. Furthermore, the sensitization with non-tuberculous mycobacteria (NTM) was also interfering the diagnosis relying in PPDs, suggesting the need of using DIVA antigens under this epidemiological condition, whether or not the BCG vaccine is administered in cattle, in order to improve the accuracy of bTB diagnosis in central Chile